Plants under herbivore attack emit volatile organic compounds (VOCs) that can serve as foraging cues for natural enemies. Adult females of Lepidoptera, when foraging for host plants to deposit eggs, are commonly repelled by herbivore-induced VOCs, probably to avoid competition and natural enemies. Their larval stages, on the other hand, have been shown to be attracted to inducible VOCs. We speculate that this contradicting behavior of lepidopteran larvae is due to a need to quickly find a new suitable host plant if they have fallen to the ground. However, once they are on a plant they might avoid the sites with fresh damage to limit competition and risk of cannibalism by conspecifics, as well as exposure to natural enemies. To test this we studied the effect of herbivore-induced VOCs on the attraction of larvae of the moth Spodopteralittoralis and on their feeding behavior. The experiments further considered the importance of previous feeding experience on the responses of the larvae. It was confirmed that herbivore-induced VOCs emitted by maize plants are attractive to the larvae, but exposure to the volatiles decreased the growth rate of caterpillars at early developmental stages. Larvae that had fed on maize previously were more attracted by VOCs of induced maize than larvae that had fed on artificial diet. At relatively high concentrations synthetic green leaf volatiles, indicative of fresh damage, also negatively affected the growth rate of caterpillars, but not at low concentrations. In all cases, feeding by the later stages of the larvae was not affected by the VOCs. The results are discussed in the context of larval foraging behavior under natural conditions, where there may be a trade-off between using available host plant signals and avoiding competitors and natural enemies.

Aqueous and organic (hexane, chloroform, and methanol) extracts of siliquae, stems and leaves, and seeds of Cleome arabica L. (Brassicales: Capparidaceae) were evaluated in the laboratory for their antifeeding and insecticidal effect on larvae of the cotton leafworm, Spodopteralittoralis (Boisduval) (Lepidoptera, Noctuidae), using a leaf dipping bioassay with castor bean, Ricinus communis L. (Malpighiales: Euphorbiaceae), leaf discs. The polar extracts caused significant mortality. At the highest dose, C. arabica extracts exhibited significant antifeeding and phagostimulating activities against S. littoralis larvae. Under no-choice conditions, the methanol extract of siliquae was the most active, and the antifeedant index calculated over 24 hr for 3rd instar larvae varied significantly from 16 to 37%. Using nutritional indices, it was established that there was a significant decrease in growth rate concomitant with a reduction in consumption. These results suggest the presence of anti-feeding and/or toxic substances in the extracts that may be useful in developing bio-insecticides based on C. arabica extracts for use in integrated pest management of leafworm and other agricultural pests. PMID:23906290

The Spodopteralittoralis multiple nucleopolyhedrovirus (SpliMNPV), a pathogen of the Egyptian cotton leaf worm Spodopteralittoralis, was subjected to sequencing of its entire DNA genome and bioassay analysis comparing its virulence to that of other baculoviruses. The annotated SpliMNPV genome of...

The genus Spodoptera comprises 31 species, 4 of which are listed as quarantine pests for the European Union: Spodoptera eridania (Cramer), Spodoptera frugiperda (Smith), Spodopteralittoralis (Boisduval), and Spodoptera litura (F.). In international trade, the earlier life stages (eggs and larvae) are being intercepted at point of inspection most frequently, challenging the possibilities of morphological identification. To realize a rapid and reliable identification for all stages, we developed and validated four simplex real-time polymerase chain reaction identification tests based on the mitochondrial cytochrome b gene using dual-labeled hydrolysis probes. Method validation on dilutions of extracted DNA of the target organisms showed that low levels of template (up to 0.2-100 pg) can reliably be identified. No cross-reactivity was observed with 14 nontarget Spodoptera and 5 non-Spodoptera species in the specific Spodoptera tests. The tests showed to be repeatable, reproducible (both 100%), and robust. The new Spodoptera tests have proven to be suitable tools for routine identification of all life stages of S. eridania, S. frugiperda, S. littoralis, and S. litura.

Plant lectins have received a lot of attention because of their insecticidal properties. When orally administered in artificial diet or in transgenic plants, lectins provoke a wide range of detrimental effects, including alteration of the digestive enzyme machinery, fecundity drop, reduced feeding, changes in oviposition behavior, growth and development inhibition and mortality. Although many studies reported the entomotoxicity of lectins, only a few of them investigated the mode of action by which lectins exert toxicity. In the present paper we have studied for the first time the insecticidal potential of the plant lectin from Hippeastrum hybrid (Amaryllis) (HHA) bulbs against the larvae of the cotton leafworm (Spodopteralittoralis). Bioassays on neonate larvae showed that this mannose-specific lectin affected larval growth, causing a development retardation and larval weight decrease. Using primary cell cultures from S. littoralis midguts and confocal microscopy we have elucidated FITC-HHA binding and internalization mechanisms. We found that HHA did not exert a toxic effect on S. littoralis midgut cells, but HHA interaction with the brush border of midgut cells interfered with normal nutrient absorption in the S. littoralis midgut, thereby affecting normal larval growth in vivo. This study thus confirms the potential of mannose-specific lectins as pest control agents and sheds light on the mechanism underlying lectin entomotoxicity.

Allelochemicals play important roles in the plant defense against herbivorous insects. They act as feeding deterrents, interfere with digestion and nutrient absorption, and cause production of potentially dangerous oxidative radicals. This study demonstrates that the distributions of oxidative radicals and of the antioxidant enzymes that eliminate them are compartmentalized in the digestive tract of Spodopteralittoralis larvae. Feeding on diets supplemented with the tannic acid (TA), alpha-solanine, and demissidine, respectively, did not affect the rate of food passage through the digestive tract of larvae but 1.25, 2.5, and 5% TA evoked a strong oxidative response. The amount of the superoxide anion in the foregut tissue and content increased up to 70-fold and the titer of total peroxides in the foregut content about 3-fold. This oxidative stress was associated with enhanced carbonyl content in the foregut tissue proteins, indicative of certain tissue deterioration. Extensive foregut damage was probably prevented by elevated activity of the glutathione S-transferase peroxidase. A complex antioxidant response was elicited in the midgut. The activities of superoxide dismutase and catalase increased significantly in the midgut tissue and content, and the activity of ascorbate peroxidase rose in the midgut tissue. The enzymes apparently eliminated oxidative radicals passing to midgut from the foregut with the food bolus and thereby prevented carbonylation of the midgut proteins. We postulate that the generation of oxidative radicals in the foregut and the induction of antioxidant defense in the midgut are controlled processes and that their compartmentalization is an important functional feature of the digestive tract. The glycoalkaloid alpha-solanine and the aglycone demissidine applied at 0.05 and 0.1% concentrations had no effect on any of the examined parameters.

Chemical senses are crucial for all organisms to detect various environmental information. Different protein families, expressed in chemosensory organs, are involved in the detection of this information, such as odorant-binding proteins, olfactory and gustatory receptors, and ionotropic receptors. We recently reported an Expressed Sequence Tag (EST) approach on male antennae of the noctuid moth, Spodopteralittoralis, with which we could identify a large array of chemosensory genes in a species for which no genomic data are available. Here we describe a complementary EST project on female antennae in the same species. 18,342 ESTs were sequenced and their assembly with our previous male ESTs led to a total of 13,685 unigenes, greatly improving our description of the S. littoralis antennal transcriptome. Gene ontology comparison between male and female data suggested a similar complexity of antennae of both sexes. Focusing on chemosensation, we identified 26 odorant-binding proteins, 36 olfactory and 5 gustatory receptors, expressed in the antennae of S. littoralis. One of the newly identified gustatory receptors appeared as female-enriched. Together with its atypical tissue-distribution, this suggests a role in oviposition. The compilation of male and female antennal ESTs represents a valuable resource for exploring the mechanisms of olfaction in S. littoralis. PMID:22904672

A crude acetone extract and oil of ripe fruits from Melia azedarach L. were evaluated against the 2nd and 4th instar larvae of Spodopteralittoralis (Boisd.) (Lepidoptera: Noctuidae). Both oil and extract exhibited highly significant growth inhibition at all concentrations tested, while the oil of M. azedarach recorded higher insecticidal activity against both instars than the crude extract. GC-MS analysis of the oil revealed the presence of linoleic acid methyl ester, oleic acid methyl ester, and free oleic acid as the main components in addition to hexadecanol, palmitic acid, methyl esters of stearic acid and myristic acid. Fatty acids and their esters were not only the main constituents of essential oil from the ripe fruits of M. azedarach, but also mainly responsible for the insecticidal and growth inhibition activity against S. littoralis.

Background Plant Ca2+ signals are involved in a wide array of intracellular signaling pathways after pest invasion. Ca2+-binding sensory proteins such as Ca2+-dependent protein kinases (CPKs) have been predicted to mediate the signaling following Ca2+ influx after insect herbivory. However, until now this prediction was not testable. Results To investigate the roles CPKs play in a herbivore response-signaling pathway, we screened the characteristics of Arabidopsis CPK mutants damaged by a feeding generalist herbivore, Spodopteralittoralis. Following insect attack, the cpk3 and cpk13 mutants showed lower transcript levels of plant defensin gene PDF1.2 compared to wild-type plants. The CPK cascade was not directly linked to the herbivory-induced signaling pathways that were mediated by defense-related phytohormones such as jasmonic acid and ethylene. CPK3 was also suggested to be involved in a negative feedback regulation of the cytosolic Ca2+ levels after herbivory and wounding damage. In vitro kinase assays of CPK3 protein with a suite of substrates demonstrated that the protein phosphorylates transcription factors (including ERF1, HsfB2a and CZF1/ZFAR1) in the presence of Ca2+. CPK13 strongly phosphorylated only HsfB2a, irrespective of the presence of Ca2+. Furthermore, in vivo agroinfiltration assays showed that CPK3-or CPK13-derived phosphorylation of a heat shock factor (HsfB2a) promotes PDF1.2 transcriptional activation in the defense response. Conclusions These results reveal the involvement of two Arabidopsis CPKs (CPK3 and CPK13) in the herbivory-induced signaling network via HsfB2a-mediated regulation of the defense-related transcriptional machinery. This cascade is not involved in the phytohormone-related signaling pathways, but rather directly impacts transcription factors for defense responses. PMID:20504319

Phytophagous insects use blends of volatiles released from plants to select hosts for feeding and oviposition. To behaviorally analyze complex blends, we need efficient and selective methods for elucidating neuron types, their ligands, and specificity. Gas chromatography-combined single sensillum recordings (GC-SSRs) from antennal olfactory sensilla of female moth, Spodopteralittoralis revealed 38 physiologically active peaks in the headspace volatile blends from both larvae-damaged cotton plants and lilac flowers. Using GC-combined mass spectrometry, 9 new physiologically active compounds were identified from damaged cotton and 11 from lilac compared with earlier electrophysiological studies using antennae of female S. littoralis. We characterized 14 novel classes of olfactory sensory neurons (OSNs). Among these, we found the first 2 ligands for a frequent type of short trichoid sensillum, for which no ligands were identified earlier. By using GC-SSR, a substantial increase in functional classes of OSNs and active compounds, 40% and 34% more, respectively, compared with recent studies using GC-electroantennogram or SSR using single compounds was detected. Compared with the estimated number of corresponding antennal olfactory receptors, the OSN classes now correspond to 83% of a likely maximum. The many specialist OSNs observed may facilitate behavioral confirmation of key plant volatiles in blends.

Four local Bacillus thuringiensis (Bt) isolates that had been serologically identified as Bt var. kurstaki (Btk2, Btk3, and Btk66) and Bt var. mexicanensis (Btm27), in addition to two reference strains (4D20 and 4AC1), were laboratory assayed as microbial control agents against the Egyptian cotton leafworm Spodopteralittoralis (Boisd.). Polymerase chain reaction (PCR) amplification analysis revealed that each of the six experimental strains carries, at least, a cry1 type gene which expresses a protein toxin active against lepidopterous insects. Additionally, PCR amplification results demonstrated that 4D20 and Btk66 contain the Lepidoptera- and Diptera-active cry2 type gene and that Btk66 contains Coleoptera-active cry7 and cry8 genes. Among the six strains, Btk66 and Btm27 were the most promising microbial control agents against S. littoralis. The present findings were the first to report that Btm27 (classified as B. thuringiensis var. mexicanensis) is a very potent microbial control agent against S. littoralis-tested larvae. For more characterization of these two isolates, the sspO gene was investigated as a molecular chronometer. The DNA sequencing results proved that Btk66 and Btm27 carry sspO open reading frames with identical nucleotide sequences, suggesting a strong phylogenetic relationship between the two strains.

The cotton leafworm, Spodopteralittoralis Boisduval (Lepidoptera: Noctuidae), is one of the most devastating pests of crops worldwide. Several types of treatments have been used against this pest, but many of them failed because of the rapid development of genetic resistance in the different insect populations. G protein coupled receptors have vital functions in most organisms, including insects; thus, they are appealing targets for species-specific pest control strategies. Among the insect G protein coupled receptors, the diuretic hormone receptors have several key roles in development and metabolism, but their importance in vivo and their potential role as targets of novel pest control strategies are largely unexplored. With the goal of using DHR genes as targets to control S. littoralis, we cloned a corticotropin-releasing factor-like binding receptor in this species and expressed the corresponding dsRNA in tobacco plants to knock down the receptor activity in vivo through RNA interference. We also expressed the receptor in mammalian cells to study its signaling pathways. The results indicate that this diuretic hormone receptor gene has vital roles in S. littoralis and represents an excellent molecular target to protect agriculturallyimportant plants from this pest. PMID:25368043

Chlorophylls, the most prominent natural pigments, are part of the daily diet of herbivorous insects. The spectrum of ingested and digested chlorophyll metabolites compares well to the pattern of early chlorophyll-degradation products in senescent plants. Intact chlorophyll is rapidly degraded by proteins in the front- and midgut. Unlike plants, insects convert both chlorophyll a and b into the corresponding catabolites. MALDI-TOF/MS imaging allowed monitoring the distribution of the chlorophyll catabolites along the gut of Spodopteralittoralis larvae. The chlorophyll degradation in the fore- and mid-gut is strongly pH dependent, and requires alkaline conditions. Using LC-MS/MS analysis we identified a lipocalin-type protein in the intestinal fluid of S. littoralis homolog to the chlorophyllide a binding protein from Bombyx mori. Widefield and high-resolution autofluorescence microscopy revealed that the brush border membranes are covered with the chlorophyllide binding protein tightly bound via its GPI-anchor to the gut membrane. A function in defense against gut microbes is discussed.

The moth Spodopteralittoralis, is a major pest of agriculture whose olfactory system is tuned to odorants emitted by host plants and conspecifics. As in other insects, the paired mushroom bodies are thought to play pivotal roles in behaviors that are elicited by contextual and multisensory signals, amongst which those of specific odors dominate. Compared with species that have elaborate behavioral repertoires, such as the honey bee Apis mellifera or the cockroach Periplaneta americana, the mushroom bodies of S. littoralis were originally viewed as having a simple cellular organization. This has been since challenged by observations of putative transmitters and neuromodulators. As revealed by immunocytology, the spodopteran mushroom bodies like those of other taxa, are subdivided longitudinally into discrete neuropil domains. Such divisions are further supported by the present study, which also demonstrates discrete affinities to different mushroom body neuropils by antibodies raised against two putative transmitters, glutamate and γ-aminobutyric acid, and against three putative neuromodulatory substances: serotonin, A-type allatostatin, and tachykinin-related peptides. The results suggest that in addition to longitudinal divisions of the lobes, circuits in the calyces and lobes are likely to be independently modulated. PMID:18406668

Spodopteralittoralis, one of the most serious and destructive agricultural pests in the world, is very susceptible to Vip3 toxin. In order to develop a new efficient bioinsecticide and to prevent the development of resistance by the target pest, insecticidal activity of biosurfactant produced by Bacillus amyloliquefaciens AG1 was evaluated against S. littoralis. Bioassays revealed the susceptibility of the first instar larvae of this pest to AG1 biosurfactant with an LC50 of 245ng/cm(2). Moreover, the histopathology examination of the larval midgut treated by AG1 biosurfactant showed vacuolization, necrosis and disintegration of the basement membrane. Binding experiments revealed that the AG1 biosurfactant recognized three putative receptors located in the brush border membrane vesicles of S. littoralis with sizes of 91, 72 and 64kDa. Competition assays using biotinylated metabolites indicated that AG1 biosurfactant and Vip3Aa16 toxin did not compete for the same S. littoralis receptors. When combined, AG1 biosurfactant and Vip3Aa16 showed an additive effect against S. littoralis larvae. These findings suggested that B. amyloliquefaciens AG1 biosurfactant could be a promising biocontrol agent to eradicate S. littoralis and to prevent resistance development by this pest.

SPB1 is a Bacillus subtilis strain producing a lipopeptide biosurfactant. The insecticidal activity of this biosurfactant was evaluated against the Egyptian cotton leaf worm (Spodopteralittoralis). It displayed toxicity with an LC(50) of 251 ng/cm(2). The histopathological changes occurred in the larval midgut of S. littoralis treated with B. subtilis SPB1 biosurfactant were vesicle formation in the apical region, cellular vacuolization and destruction of epithelial cells and their boundaries. Ligand-blotting experiments with S. littoralis brush border membrane vesicles showed binding of SPB1 biosurfactant to a protein of 45 kDa corresponding to its putative receptor. The latter differs in molecular size from those recognized by Bacillus thuringiensis Vip3A and Cry1C toxins, commonly known by their activity against S. littoralis. This result wires the application of B. subtilis biosurfactant for effective control of S. littoralis larvae, particularly in the cases where S. littoralis will develop resistance against B. thuringiensis toxins.

Background Biotic stress induced by various herbivores and pathogens invokes plant responses involving different defense mechanisms. However, we do not know whether different biotic stresses share a common response or which signaling pathways are involved in responses to different biotic stresses. We investigated the common and specific responses of Arabidopsis thaliana to three biotic stress agents: Spodopteralittoralis, Myzus persicae, and the pathogen Pseudomonas syringae. Methodology/Principal Findings We used electrophysiology to determine the plasma membrane potential (Vm) and we performed a gene microarray transcriptome analysis on Arabidopsis upon either herbivory or bacterial infection. Vm depolarization was induced by insect attack; however, the response was much more rapid to S. littoralis (30 min −2 h) than to M. persicae (4–6 h). M. persicae differentially regulated almost 10-fold more genes than by S. littoralis with an opposite regulation. M. persicae modulated genes involved in flavonoid, fatty acid, hormone, drug transport and chitin metabolism. S. littoralis regulated responses to heat, transcription and ion transport. The latest Vm depolarization (16 h) was found for P. syringae. The pathogen regulated responses to salicylate, jasmonate and to microorganisms. Despite this late response, the number of genes differentially regulated by P. syringae was closer to those regulated by S. littoralis than by M. persicae. Conclusions/Significance Arabidopsis plasma membranes respond with a Vm depolarization at times depending on the nature of biotic attack which allow setting a time point for comparative genome-wide analysis. A clear relationship between Vm depolarization and gene expression was found. At Vm depolarization timing, M. persicae regulates a wider array of Arabidopsis genes with a clear and distinct regulation than S. littoralis. An almost completely opposite regulation was observed between the aphid and the pathogen, with the former

Natural odors are often complex mixtures of different compounds. These mixtures can be perceived to have qualities that are different from their components. Moreover, components can be difficult to distinguish within a blend, even if those components are identifiable when presented individually. Thus, odor components can interact along the olfactory pathway in a nonlinear fashion such that the mixture is not perceived simply as the sum of its components. Here we investigated odor-evoked changes in Ca2+ concentration to binary blends of plant-related substances in individually identified glomeruli in the moth Spodopteralittoralis. We used a wide range of blend ratios and a range of concentrations below the level at which glomerular responses become saturated. We found no statistically significant cases where the mixture response was greater than both component responses at the same total concentration (synergistic interactions) and no statistically significant cases where the mixture response was less than either component presented individually (suppressive interactions). Therefore, we conclude that, for the plant mixtures studied, information of their components is preserved in the neural representations encoded at the first stage of olfactory processing in this moth species.

The gut microbiota is of crucial importance for the host with considerable metabolic activity. Although great efforts have been made toward characterizing microbial diversity, measuring components' metabolic activity surprisingly hasn't kept pace. Here we combined pyrosequencing of amplified 16S rRNA genes with in vivo stable isotope probing (Pyro-SIP) to unmask metabolically active bacteria in the gut of cotton leafworm (Spodopteralittoralis), a polyphagous insect herbivore that consumes large amounts of plant material in a short time, liberating abundant glucose in the alimentary canal as a most important carbon and energy source for both host and active gut bacteria. With (13)C glucose as the trophic link, Pyro-SIP revealed that a relatively simple but distinctive gut microbiota co-developed with the host, both metabolic activity and composition shifting throughout larval stages. Pantoea, Citrobacter and Clostridium were particularly active in early-instar, likely the core functional populations linked to nutritional upgrading. Enterococcus was the single predominant genus in the community, and it was essentially stable and metabolically active in the larval lifespan. Based on that Enterococci formed biofilm-like layers on the gut epithelium and that the isolated strains showed antimicrobial properties, Enterococcus may be able to establish a colonization resistance effect in the gut against potentially harmful microbes from outside. Not only does this establish the first in-depth inventory of the gut microbiota of a model organism from the mostly phytophagous Lepidoptera, but this pilot study shows that Pyro-SIP can rapidly gain insight into the gut microbiota's metabolic activity with high resolution and high precision.

The gut microbiota is of crucial importance for the host with considerable metabolic activity. Although great efforts have been made toward characterizing microbial diversity, measuring components' metabolic activity surprisingly hasn't kept pace. Here we combined pyrosequencing of amplified 16S rRNA genes with in vivo stable isotope probing (Pyro-SIP) to unmask metabolically active bacteria in the gut of cotton leafworm (Spodopteralittoralis), a polyphagous insect herbivore that consumes large amounts of plant material in a short time, liberating abundant glucose in the alimentary canal as a most important carbon and energy source for both host and active gut bacteria. With 13C glucose as the trophic link, Pyro-SIP revealed that a relatively simple but distinctive gut microbiota co-developed with the host, both metabolic activity and composition shifting throughout larval stages. Pantoea, Citrobacter and Clostridium were particularly active in early-instar, likely the core functional populations linked to nutritional upgrading. Enterococcus was the single predominant genus in the community, and it was essentially stable and metabolically active in the larval lifespan. Based on that Enterococci formed biofilm-like layers on the gut epithelium and that the isolated strains showed antimicrobial properties, Enterococcus may be able to establish a colonization resistance effect in the gut against potentially harmful microbes from outside. Not only does this establish the first in-depth inventory of the gut microbiota of a model organism from the mostly phytophagous Lepidoptera, but this pilot study shows that Pyro-SIP can rapidly gain insight into the gut microbiota's metabolic activity with high resolution and high precision. PMID:24475063

Bacillus thuringiensis subsp. aizawai strain HD133, known by its effectiveness against Spodoptera species, produces many insecticidal proteins including Cry1Ab, Cry1Ca and Cry1Da. In the present study, the insecticidal activity of Cry1Da against Spodopteralittoralis was investigated. It showed toxicity with an LC(50) of 224.4 ng/cm(2) with 95% confidence limits of (178.61-270.19) and an LC(90) of 467.77 ng/cm(2) with 95% confidence limits of (392.89-542.65). The midgut histopathology of Cry1Da fed larvae showed vesicle formation in the apical region, vacuolization and destruction of epithelial cells. Biotinylated-activated Cry1Da toxin bound protein of about 65 kDa on blots of S. littoralis brush border membrane preparations. This putative receptor differs in molecular size from those recognized by Cry1C and Vip3A which are active against this polyphagous insect. This difference in midgut receptors strongly supports the use of Cry1Da as insecticidal agent, particularly in case of Cry and/or Vip-resistance management.

Fatty acyl-CoA reductases (FARs), the enzymes that catalyse reduction of a fatty acyl-CoA to the corresponding alcohol in insect pheromone biosynthesis, are postulated to play an important role in determining the proportion of each component in the pheromone blend. For the first time, we have isolated and characterized from the Egyptian cotton leaf worm Spodopteralittoralis (Lepidoptera: Noctuidae) a FAR cDNA (Slit-FAR1), which appeared to be expressed only in the pheromone gland and was undetectable in other female tissues, such as fat body, ovaries, wings, legs or thorax. The encoded protein has been successfully expressed in a recombinant system, and the recombinant enzyme is able to produce the intermediate fatty acid alcohols of the pheromone biosynthesis of S. littoralis from the corresponding acyl-CoA precursors. The kinetic variables Km and Vmax, which have been calculated for each acyl-CoA pheromone precursor, suggest that in S. littoralis pheromone biosynthesis other biosynthetic enzymes (e.g. desaturases, acetyl transferase) should also contribute to the final ratio of components of the pheromone blend. In a phylogenetic analysis, Slit-FAR1 appeared grouped in a cluster of other FARs involved in the pheromone biosynthesis of other insects, with little or non-specificity for the natural pheromone precursors.

Circadian clocks (oscillators) regulate multiple life functions in insects. The circadian system located in the male reproductive tract of Lepidoptera is one of the best characterized peripheral oscillators in insects. Our previous research on the cotton leafworm, Spodopteralittoralis, demonstrated that this oscillator controls the rhythm of sperm release from the testis and coordinates sperm maturation in the upper vas deferens (UVD). We demonstrated previously that a protein that functions as yolk protein in females is also produced in cyst cells surrounding sperm bundles in the testis, and is released into the UVD. Here, we investigated the temporal expression of the yolk protein 2 (yp2) gene at the mRNA and protein level in the testis of S. littoralis, and inquired whether their expression is regulated by PER-based molecular oscillator. We describe a circadian rhythm of YP2 accumulation in the UVD seminal fluid, where this protein interacts with sperm in a circadian fashion. However, we also demonstrate that yp2 mRNA and YP2 protein levels within cyst cells show only a diurnal rhythm in light/dark (LD) cycles. These rhythms do not persist in constant darkness (DD), suggesting that they are non-circadian. Interestingly, the per gene mRNA and protein levels in cyst cells are rhythmic in LD but not in DD. Nevertheless, per appears to be involved in the diurnal timing of YP2 protein accumulation in cyst cells.

The bacterium Bacillus thuringiensis produces, at the vegetative stage of its growth, Vip3A proteins with activity against a broad spectrum of lepidopteran insects. The Egyptian cotton leaf worm (Spodopteralittoralis) is an important agricultural pest that is susceptible to the Vip3Aa16 protein of Bacillus thuringiensis kurstaki strain BUPM95. The midgut histopathology of Vip3Aa fed larvae showed vacuolization of the cytoplasm, brush border membrane destruction, vesicle formation in the apical region and cellular disintegration. Biotinylated Vip3Aa toxin bound proteins of 55- and 100-kDa on blots of S. littoralis brush border membrane preparations. These binding proteins differ in molecular size from those recognized by Cry1C, one of the very few Cry proteins active against the polyphagous S. littoralis. This result supports the use of Vip3Aa16 proteins as insecticidal agent, especially in case of Cry-resistance management.

Moth sex pheromone communication is recognised as a long-standing model for insect olfaction studies, and a widespread knowledge has been accumulated on this subject thanks to numerous chemical, electrophysiological and behavioural studies. A key step has been the identification of candidate sex pheromone receptors, opening new routes to understanding the specificity and sensitivity of this communication system, but only few of these receptors have as yet been functionally characterised. In this context, we aim at unravelling the molecular bases of pheromone reception in the noctuid moth Spodopteralittoralis. Taking advantage of a collection of antennal-expressed sequence tags, we previously identified three fragments of candidate pheromone receptors in this species. Here, we report full-length cloning of one of these receptors, named SlitOR6. Both sequence and expression pattern analyses were consistent with its annotation as a pheromone receptor, which we further confirmed by functional characterization. Using Drosophila antennae as a heterologous expression system, we identified a single component of the pheromone blend of S. littoralis, (Z,E)-9,12-tetradecadienyl acetate, as the ligand of SlitOR6. Two strategies were employed: (i) expressing SlitOR6 in the majority of Drosophila olfactory neurons, in addition to endogenous receptors, and monitoring the responses to pheromone stimuli by electroantennography; (ii) replacing the Drosophila pheromone receptor OR67d with SlitOR6 and monitoring the response by single sensillum recordings. Results were fully congruent and responses to (Z,E)-9,12-tetradecadienyl acetate were highly specific in both heterologous systems. This approach appears to be efficient and reliable for studying moth pheromone receptors in an in vivo context.

The pre-ovipositon behavior of moths is largely dependent upon the cues that a gravid female perceives while assessing potential oviposition sites. Assessment of such sites is accomplished, at least in part, by mechanosensory and gustatory sensilla located on the ovipositor whose sensory neurons project into the terminal abdominal ganglion (TAG). Using anterograde backfill staining, confocal laser scanning microscopy, and three dimensional reconstruction, we traced and analyzed the central projections of the sensory neurons housed in the sensilla located on the ovipositor papillae and explored the neuropilar composition of the TAG in the Egyptian cotton leafworm, Spodopteralittoralis. The TAG consists of three fused neuromeres (6-8th Ner) associated with the 6-8th abdominal segments. Within the TAG, and specifically in the 8th neuromere, four unstructured neuropilar compartments are present; the dorso-ipsilateral motor neuropil (MN), the medio-ipsilateral mechanosensory neuropil (MchN), the medio-ipsilateral small gustatory neuropil (GN), and the medio-contralateral posterior ovipositor glomerulus (Og). The Og appears quite compact, with a hollow core free of terminal arborizations. The MchN is further subdivided into 4 unstructured glomeruli in the 8th neuromere, whose afferents are subsequently extended into 3 glomeruli in the 7th and 6th neuromeres. Few neurites of the Og are populated with large dense varicosities reminiscent of neurosecretory vesicles. Given that all ovipositor nerves converge into a common ganglionic center, the TAG, we assume that this ganglion may be a center for coordination of oviposition behaviors, including movements of the ovipositor during assessment of oviposition substrates and egg laying in S. littoralis.

Due to the importance of chitinolytic enzymes for insect, nematode and fungal growth, they are receiving attention concerning their development as biopesticides or chemical defense proteins in transgenic plants and as microbial biocontrol agents. Targeting chitin associated with the extracellular matrices or cell wall by insect chitinases may be an effective approach for controlling pest insects and pathogenic fungi. The ability of chitinases to attack and digest chitin in the peritrophic matrix or exoskeleton raises the possibility to use them as insect control method. In this study, an insect chitinase cDNA from cotton leaf worm (Spodopteralittoralis) has been synthesized. Transgenic maize plant system was used to improve its tolerance against insects. Insect chitinase transcripts and proteins were expressed in transgenic maize plants. The functional integrity and expression of chitinase in progenies of the transgenic plants were confirmed by insect bioassays. The bioassays using transgenic corn plants against corn borer (Sesamia cretica) revealed that ~50% of the insects reared on transgenic corn plants died, suggesting that transgenic maize plants have enhanced resistance against S. cretica.

In an attempt to increase the insecticidal effect of the delta-endotoxin crystal protein CryIC on the relatively Cry-insensitive larvae of Spodopteralittoralis, a combination of CryIC and endochitinase was used. CryIC comprising the first 756 amino acids from Bacillus thuringiensis K26-21 and endochitinase ChiAII encoded by Serratia marcescens were separately produced in Escherichia coli carrying the genes in overexpression vectors. The endochitinase on its own, even at very low concentrations (0.1 microgram/ml), perforated the larval midgut peritrophic membrane. When applied together with low concentrations of CryIC, a synergistic toxic effect was obtained. In the absence of chitinase, about 20 micrograms of CryIC per ml was required to obtain maximal reduction in larval weight, while only 3.0 micrograms of CryIC per ml caused a similar toxic effect in the presence of endochitinase. Thus, a combination of the Cry protein and an endochitinase could result in effective insect control in transgenic systems in which the Cry protein is not expressed in a crystalline form. PMID:8837413

In this study, we undertook a survey to analyze the distribution and frequency of microsatellites or Simple Sequence Repeats (SSRs) in Spodopteralittoralis multiple nucleopolyhedrovirus (SpliMNPV) genome (isolate AN–1956). Out of the 55 microsatellite motifs, identified in the SpliMNPV-AN1956 genome using in silico analysis (inclusive of mono-, di-, tri- and hexa-nucleotide repeats), 39 were found to be distributed within coding regions (cSSRs), whereas 16 were observed to lie within intergenic or noncoding regions. Among the 39 motifs located in coding regions, 21 were located in annotated functional genes whilst 18 were identified in unknown functional genes (hypothetical proteins). Among the identified motifs, trinucleotide (80%) repeats were found to be the most abundant followed by dinucleotide (13%), mononucleotide (5%) and hexanucleotide (2%) repeats. The 39 motifs located within coding regions were further validated in vitro by using PCR analysis, while the 21 motifs located within known functional genes (15 genes) were characterized using nucleotide sequencing. A comparison of the sequence analysis data of the 21 sequenced cSSRs with the published sequences is presented. Finally, the developed SSR markers of the 39 motifs were further mapped/localized onto the SpliMNPV-AN1956 genome. In conclusion, the SSR markers specific to SpliMNPV, developed in this study, could be a useful tool for the identification of isolates and analysis of genetic diversity and viral evolutionary status. PMID:27650818

Due to the importance of chitinolytic enzymes for insect, nematode and fungal growth, they are receiving attention concerning their development as biopesticides or chemical defense proteins in transgenic plants and as microbial biocontrol agents. Targeting chitin associated with the extracellular matrices or cell wall by insect chitinases may be an effective approach for controlling pest insects and pathogenic fungi. The ability of chitinases to attack and digest chitin in the peritrophic matrix or exoskeleton raises the possibility to use them as insect control method. In this study, an insect chitinase cDNA from cotton leaf worm (Spodopteralittoralis) has been synthesized. Transgenic maize plant system was used to improve its tolerance against insects. Insect chitinase transcripts and proteins were expressed in transgenic maize plants. The functional integrity and expression of chitinase in progenies of the transgenic plants were confirmed by insect bioassays. The bioassays using transgenic corn plants against corn borer (Sesamia cretica) revealed that ~50% of the insects reared on transgenic corn plants died, suggesting that transgenic maize plants have enhanced resistance against S. cretica. PMID:26658494

Microbes that live inside insects play critical roles in host nutrition, physiology, and behavior. Although Lepidoptera (butterflies and moths) are one of the most diverse insect taxa, their microbial symbionts are little-studied, particularly during metamorphosis. Here, using ribosomal tag pyrosequencing of DNA and RNA, we investigated biodiversity and activity of gut microbiotas across the holometabolous life cycle of Spodopteralittoralis, a notorious agricultural pest worldwide. Proteobacteria and Firmicutes dominate but undergo a structural “metamorphosis” in tandem with its host. Enterococcus, Pantoea and Citrobacter were abundant and active in early-instar, while Clostridia increased in late-instar. Interestingly, only enterococci persisted through metamorphosis. Female adults harbored high proportions of Enterococcus, Klebsiella and Pantoea, whereas males largely shifted to Klebsiella. Comparative functional analysis with PICRUSt indicated that early-instar larval microbiome was more enriched for genes involved in cell motility and carbohydrate metabolism, whereas in late-instar amino acid, cofactor and vitamin metabolism increased. Genes involved in energy and nucleotide metabolism were abundant in pupae. Female adult microbiome was enriched for genes relevant to energy metabolism, while an increase in the replication and repair pathway was observed in male. Understanding the metabolic activity of these herbivore-associated microbial symbionts may assist the development of novel pest-management strategies. PMID:27389097

To better understand the olfactory mechanisms in a lepidopteran pest model species, the cotton leafworm Spodopteralittoralis, we have recently established a partial transcriptome from adult antennae. Here, we completed this transcriptome using next generation sequencing technologies, namely 454 and Illumina, on both adult antennae and larval tissues, including caterpillar antennae and maxillary palps. All sequences were assembled in 77,643 contigs. Their analysis greatly enriched the repertoire of chemosensory genes in this species, with a total of 57 candidate odorant-binding and chemosensory proteins, 47 olfactory receptors, 6 gustatory receptors and 17 ionotropic receptors. Using RT-PCR, we conducted the first exhaustive comparison of olfactory gene expression between larvae and adults in a lepidopteran species. All the 127 candidate olfactory genes were profiled for expression in male and female adult antennae and in caterpillar antennae and maxillary palps. We found that caterpillars expressed a smaller set of olfactory genes than adults, with a large overlap between these two developmental stages. Two binding proteins appeared to be larvae-specific and two others were adult-specific. Interestingly, comparison between caterpillar antennae and maxillary palps revealed numerous organ-specific transcripts, suggesting the complementary involvement of these two organs in larval chemosensory detection. Adult males and females shared the same set of olfactory transcripts, except two male-specific candidate pheromone receptors, two male-specific and two female-specific odorant-binding proteins. This study identified transcripts that may be important for sex-specific or developmental stage-specific chemosensory behaviors. PMID:23565215

A sample of Spodopteralittoralis pupae were field collected from in Lebanon in November 1999. Approximately 50 healthy pupae were used to initiate a laboratory colony (stain code LEB). As a benchmark for sensitivity to insecticides the susceptible laboratory strain (LET) was used for comparison. The LET stain has been held in laboratory culture for over 10 years and originated from Israel. Three test chemicals were investigated; Spinosad (Tracer, 480 g/L SC), chlorpyrifos (Dursban 480 g/L EC) and cypermethrin (Ambush C 100 g/L EC) representing a range of different modes of actions. To estimate the sensitivity of the LEB strain relative to the LET stain laboratory bioassays, conducted on second instar larvae exposed treated tomato leaves for two days. The method used was based on IRAC (Insecticide Resistance Action Committee) insecticide susceptibly test number 7. Larvae from the LEB strain were tested in both the F1 and F2 laboratory generations. LC50 values were calculated using a Log-Probit model allowing the susceptibly of each strain to each test item to be compared. The LEB (field) strain were approximately twice as sensitive to spinosad compared with the LET (laboratory) strain. The LEB strain was 6-10 times less sensitive to chlorpyrifos which was within the range expected. However the LEB stain was 250-360 times less sensitive to cypermethrin compared with the LET strain, indicating a high level of resistance to pyrethroids in the field population. This kind of relationship gave a strong indication for negative cross resistance between spinosad and cypermethrin. It was concluded that spinosad had potential as a resistance management tool due to its novel mode of action and negative cross resistance with pyrethroids.

Serine protease inhibitors (PIs) have been described in many plant species and are universal throughout the plant kingdom, where trypsin inhibitors is the most common type. In the present study, trypsin and chymotrypsin inhibitory activity was detected in the seed flour extracts of 13 selected cultivars/accessions of cowpea. Two cowpea cultivars, Cream7 and Buff, were found to have higher trypsin and chymotrypsin inhibitory potential compared to other tested cultivars for which they have been selected for further purification studies using ammonium sulfate fractionation and DEAE-Sephadex A-25 column. Cream7-purified proteins showed two bands on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) corresponding to molecular mass of 17.10 and 14.90 kDa, while the purified protein from Buff cultivar showed a single band corresponding mass of 16.50 kDa. The purified inhibitors were stable at temperature below 60°C and were active at wide range of pH from 2 to 12. The kinetic analysis revealed noncompetitive type of inhibition for both inhibitors against both enzymes. The inhibitor constant (Ki ) values suggested high affinity between inhibitors and enzymes. Purified inhibitors were found to have deep and negative effects on the mean larval weight, larval mortality, pupation, and mean pupal weight of Spodopteralittoralis, where Buff PI was more effective than Cream7 PI. It may be concluded that cowpea PI gene(s) could be potential insect control protein for future studies in developing insect-resistant transgenic plants.

The present work was conducted to evaluate the efficiency of two essential oils ((Flax or "Linseed" and Sesame ), five volatile plant oils ( Camphor, Red basil, Rose, Menthol and Clove ), four pesticides (Methoxyfenozide; Permethrin; Profenofos and Spinosad) and their mixtures on the cotton leaf-worm Spodopteralittoralis (Boisd.). This study was also devoted to minimize the usage of conventional insecticides, reduce the environmental pollution, and protect human-beings and domestic animals from hazards due to pesticides applications. In the meantime, the delayed effect of these tested plant oils on the developing immatures and moths of the cotton leaf-worm, Spodopteralittoralis was determined. Most of the evaluated plant oils were found to have an insecticidal effect on the 4th instar larvae of the cotton leaf-worm, S. littoralis. Both the essential Sesame oil and Clove volatile oil showed rather weak toxic effect corresponding to the same concentrations and periods of the bioassay tests. Comparing the toxicity of the tested plant oils, it was affirmed that both Rose and Red basil volatile oils were the highest efficient natural phytocompounds against the treated larvae and alternatively ranked either the 1st and/or the 2nd rank, throughout the different periods of the bioassay tests, followed by the other three tested oils which were more or less efficient phytocompounds. According to the toxicity index, all the tested oils were less toxic than the superior Red basil volatile oil after 48 h. post treatment followed by the gradual decrease in toxicity of Rose, Flax and Menthol, respectively. The development of the treated 4th larval instar was blocked due to treatment with the tested plant oils. With no exception, all the efficiently tested essential and/or volatile oils acted principally as Insect Growth Inhibitors (IGIs) rather than antifeedants causing disruption of the insect development, abnormal larvae, pupae and adults that were lead finally to death. The

The leaf extract and its fractions of Peganum harmala L. have shown pronounced mortal effect, decreased percent pupation and adult emergence of the cotton leaf worm, Spodopteralittoralis Boisd. The third instar larvae fed for two days on treated leaves were more susceptible to plant extract and its ethyl acetate and chloroform fractions. The active lowest concentration (5%) of the leaf fractions of P. harmala showed significant effect on the percentage of emerged adult parasitoids, Microplitis rufiventris Kok. GC/MS analysis showed the major constituent in ethyl acetate fraction was (23S) ethylcholest-5-en-3 beta-ol (28.04%) while those of chloroform fraction were hydroxyfuranocoumarin (Bergaptol) (15.68%), piperidinone (12.08%), thymol (11.82%), phosphoric acid, tributyl ester (9.80%) and trimethyl-nonenol (9.66%). The medicinal plant P. harmala could be carefully applied in integrated pest management due to its strong effect on cotton leaf worm pest.

Bilins in complex with specific proteins play key roles in many forms of life. Biliproteins have also been isolated from insects; however, structural details are rare and possible functions largely unknown. Recently, we identified a high-molecular weight biliprotein from a moth, Cerura vinula, as an arylphorin-type hexameric storage protein linked to a novel farnesyl biliverdin IXα; its unusual structure suggests formation by cleavage of mitochondrial heme A. In the present study of another moth, Spodopteralittoralis, we isolated two different biliproteins. These proteins were identified as a very high-density lipoprotein (VHDL) and as vitellin, respectively, by mass spectrometric sequencing. Both proteins are associated with three different farnesyl biliverdins IXα: the one bilin isolated from C. vinula and two new structurally closely related bilins, supposed to be intermediates of heme A degradation. The different bilin composition of the two biliproteins suggests that the presumed oxidations at the farnesyl side-chain take place mainly during egg development. The egg bilins are supposedly transferred from hemolymph VHDL to vitellin in the female. Both biliproteins show strong induced circular dichroism activity compatible with a predominance of the M-conformation of the bilins. This conformation is opposite to that of the arylphorin-type biliprotein from C. vinula. Electron microscopy of the VHDL-type biliprotein from S. littoralis provided a preliminary view of its structure as a homodimer and confirmed the biochemically determined molecular mass of ∼350 kDa. Further, images of S. littoralis hexamerins revealed a 2 × 3 construction identical to that known from the hexamerin from C. vinula.

Infection of Spodopteralittoralis SL2 cells with the baculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV) results in apoptosis and low yields of viral progeny, in contrast to infection with S. littoralis nucleopolyhedrovirus (SlNPV). By cotransfecting SL2 cells with AcMNPV genomic DNA and a cosmid library representing the complete SlNPV genome, we were able to rescue AcMNPV replication and to isolate recombinant virus vAcSL2, which replicated efficiently in SL2 cells. Moreover, vAcSL2 showed enhanced infectivity for S. littoralis larvae compared to AcMNPV. The genome of vAcSL2 carried a 519-bp insert fragment that increased the distance between the TATA element and the transcriptional initiation site (CAGT) of immediate-early gene ie0. This finding correlated with low steady-state levels of IE0 and higher steady-state levels of IE1 (the product of the ie1 gene, a major AcMNPV transactivator, and a multifunctional protein) than of IE0. Mutagenesis of the ie0 promoter locus by insertion of the chloramphenical acetyltransferase (cat) gene yielded a new recombinant AcMNPV with replication properties identical to those of vAcSL2. Thus, the analysis indicated that increasing the steady-state levels of IE1 relative to IE0 should enable AcMNPV replication in SL2 cells. This suggestion was confirmed by constructing a recombinant AcMNPV bearing an extra copy of the ie1 gene under the control of the Drosophila hsp70 promoter. These results suggest that IE0 plays a role in the regulation of AcMNPV infection and show, for the first time, that significant improvement in the ability of AcMNPV to replicate in a poorly permissive cell line and organism can be achieved by increasing the expression of the main multiple functional protein, IE1.

The alkaline gut of Lepidopterans plays a crucial role in shaping communities of bacteria. Enterococcus mundtii has emerged as one of the predominant gut microorganisms in the gastrointestinal tract of the major agricultural pest, Spodopteralittoralis. Therefore, it was selected as a model bacterium to study its adaptation to harsh alkaline gut conditions in its host insect throughout different stages of development (larvae, pupae, adults, and eggs). To date, the mechanism of bacterial survival in insects' intestinal tract has been unknown. Therefore, we have engineered a GFP-tagged species of bacteria, E. mundtii, to track how it colonizes the intestine of S. littoralis. Three promoters of different strengths were used to control the expression of GFP in E. mundtii. The promoter ermB was the most effective, exhibiting the highest GFP fluorescence intensity, and hence was chosen as our main construct. Our data show that the engineered fluorescent bacteria survived and proliferated in the intestinal tract of the insect at all life stages for up to the second generation following ingestion. PMID:27379058

Insecticides have long been used as the main method in limiting agricultural pests, but their widespread use has resulted in environmental pollution, development of resistances, and biodiversity reduction. The effects of insecticides at low residual doses on both the targeted crop pest species and beneficial insects have become a major concern. In particular, these low doses can induce unexpected positive (hormetic) effects on pest insects, such as surges in population growth exceeding what would have been observed without pesticide application. Methomyl and chlorpyrifos are two insecticides commonly used to control the population levels of the cotton leafworm Spodopteralittoralis, a major pest moth. The aim of the present study was to examine the effects of sublethal doses of these two pesticides, known to present a residual activity and persistence in the environment, on the moth physiology. Using a metabolomic approach, we showed that sublethal doses of methomyl and chlorpyrifos have a systemic effect on the treated insects. We also demonstrated a behavioral disruption of S. littoralis larvae exposed to sublethal doses of methomyl, whereas no effects were observed for the same doses of chlorpyrifos. Interestingly, we highlighted that sublethal doses of both pesticides did not induce a change in acetylcholinesterase activity in head of exposed larvae.

EPR spectroscopy was used to investigate the cytochrome P-450-dependent steroid hydroxylase ecdysone 20-mono-oxygenase of the cotton leafworm (Spodopteralittoralis) and the redox centres associated with membranes from the fat-body mitochondrial fraction. Intense features at g = 2.42, 2.25 and 1.92 from oxidized mitochondrial membranes have been assigned to the low-spin haem form of ferricytochrome P-450, probably of ecdysone 20-mono-oxygenase. High-spin cytochrome P-450 (substrate-bound) was tentatively assigned to a signal at g = 8.0, which was detectable from membranes as prepared. An EPR signal characteristic of a [2Fe-2S] cluster detected from the soluble mitochondrial matrix fraction has been shown to be distinct from the signals associated with mitochondrial NADH dehydrogenase and succinate dehydrogenase, and has therefore been attributed to a ferredoxin. We conclude that the S. littoralis fat-body mitochondrial electron-transport system involved in steroid 20-hydroxylation comprises both ferredoxin and cytochrome P-450 components, and thus resembles the enzyme systems of adrenocortical mitochondria. EPR signals characteristic of the respiratory chain were also observed from fat-body mitochondria and assigned to the iron-sulphur clusters associated with Complex I (Centres N1, N2), Complex II (Centres S1, S3), Complex III (the Rieske centre), and the copper centre of Complex IV, demonstrating similarities to mammalian mitochondria. The reduced membrane fraction also yielded a major resonance at g = 2.09 and 1.88 characteristic of the [4Fe-4S] cluster of electron-transferring flavoprotein: ubiquinone oxidoreductase. As the fat-body is the major metabolic organ of insects, this protein is presumably required for the beta-oxidation of fatty acids in mitochondria. High-spin haem signals in the low-field region of spectra also demonstrated that the mitochondrial fraction contains relatively high concentrations of catalase. PMID:7741702

Entomopathogenic nematodes are generally considered beneficial organisms. However, they can affect beneficial insects such as parasitoids. The interaction between the entomopathogenic nematodes Heterorhabditis bacteriophora Poinar (Rhabditida: Heterorhabditidae) and Steinernema carpocapsae Weiser, and the parasitoid Microplitis rufiventris Kokujev (Hymenoptera: Braconidae) was investigated in the laboratory. In non-parasitized hosts, Spodopteralittoralis Boisduval (Lepidoptera: Noctuidae) larvae exposed to H. bacteriophora showed a higher percent mortality than those exposed to S. carpocapsae. Both nematodes were able to invade and propagate in non-parasitized S. littoralis larvae and those parasitized by M. rufiventris. Both nematode species reproduced in Microplitis-parasitized hosts, but there was a higher number of nematodes in non-parasitized larvae. S. carpocapsae yielded higher numbers of infective juveniles than H. bacteriophora. Generally, the number of nematodes harvested increased as their host's size increased. The interaction between the nematodes and parasitoid favored the nematodes when the nematodes were inoculated during the parasitoid egg stage or the young parasitoid larvae, thus giving the nematodes a better chance to grow and reproduce, resulting in the death of the parasitoid larvae. Conversely, when the nematodes were inoculated during the late larval instar of the parasitoid, the competition partially favored the wasp, thus giving approximately 50% of the wasps a better chance to develop, emerge, and reproduce, providing evidence that both nematodes and wasps could reproduce in the same host. Egg maturation of female wasps derived from nematode-infected hosts was not significantly different than those from control hosts. The combined application of nematodes and parasitoids may be beneficial if the detrimental effects of the nematodes on the parasitoid could be avoided by precisely timing the application strategies. It is clear that

In Spodopteralittoralis, testicular sperm release occurs in a daily rhythm, which is controlled by endogenous circadian oscillator located in the male reproductive system. Although this rhythm is essential for male fertility, factors that initiate and maintain daily sperm release are not understood. In this study, we investigated a modulatory role for ecdysteroids in the sperm release rhythm and identified the source of ecdysteroids in adult males. We found that the onset of sperm release occurs two days pre-eclosion and coincides with a significant decrease in haemolymph ecdysteroids levels. 20-HE injection into the pupae prior to the first sperm release delayed its initiation and disrupted the developing rhythm in a dose dependent manner. 20-HE injection into adults depressed the number of sperm bundles leaving the testes. A day before the initial sperm release, ecdysteroid levels in the haemolymph and testes begin to oscillate in a circadian fashion. Ecdysteroid rhythms continue throughout imaginal life and correlate with the rhythm of sperm release. In each cycle, testicular sperm release coincides with a trough in testicular ecdysteroid concentration. Rhythmic changes in ecdysteroid levels are regulated by an endogenous circadian oscillator that continues to function in decapitated males. The generation of a complete cycle of ecdysteroid release by testes cultured in vitro indicates that this oscillator is located in the gonads. The haemolymph ecdysteroid levels are significantly lower and arrhythmic in males with removed testes, indicating that the testes are an important ecdysteroid source that may contribute to oscillations in haemolymph ecdysteroid levels.

The flower extract of Chrysanthemum coronarium L. and their fractions have shown insecticidal effect on the cotton leaf worm Spodopteralittoralis. The third instar larvae fed for two days on treated leaves were more susceptible to plant extracts and to their ethyl acetate and chloroform fractions. The active lowest concentration (5%) of the flower fractions showed no significant effect on the percent reduction of emerged adult parasitoids, Microplitis rufiventris Kok. GC/MS analysis revealed that the major constituents in ethyl acetate fraction were 3-dihydro-methylene-2- (3H) furanone (17.8%), jasmolin I (15.6%), carveol 1 (13.6%), phosphoric acid, tributyl ester (11.4%) and cinerin II (11.1%), while those of chloroform fraction were 5-hydroxy-3 methyl-1H-pyrazole (42.7%) and carveol 1(24.8%). The medicinal plant C. coronarium seems to be a promising plant for application in integrated pest management due to its safety to the surrounding environment.

The in vitro secretion of ecdysteroids from the prothoracic glands of last instar larvae of Spodopteralittoralis was detected and analysed by HPLC-RIA. The primary product was identified as 3-dehydroecdysone (approximately 82%), with lesser amounts of ecdysone (approximately 18%). Interconversion of ecdysone and 3-dehydroecdysone by prothoracic glands was not detectable. 3-Dehydroecdysone 3 beta-reductase activity was demonstrated in the haemolymph. Ecdysone, the endproduct, was characterised by reverse-phase and adsorption HPLC, chemical transformation into ecdysone 2, 3-acetonide, and mass spectrometry. The conditions for optimal activity were determined. The enzyme requires NADPH or NADH as cofactor and Km values for NADPH and NADH were determined to be 0.94 microM, and 22.8 microM, respectively. Investigation of the kinetic properties of the enzyme, using either NADPH or NADH as cofactor, revealed that it exhibits maximal activity at low 3-dehydroecdysone substrate concentrations, with a drastic inhibition of activity at higher concentrations (> 5 microM). The results suggest that the 3-dehydroecdysone 3 beta-reductase has a high-affinity (low Km) binding site for 3-dehydroecdysone substrate, together with a lower-affinity inhibition site. The 3 beta-reductase enzyme was purified to homogeneity using a combination of poly(ethylene glycol) 6000 precipitation and successive FPLC fractionation on Mono-Q, phenyl Superose (twice), and hydroxyapatite columns. The native enzyme was shown to be a monomer with molecular mass of 36 kDa by SDS/PAGE and gel-filtration chromatography. Furthermore, the activity of the enzyme during the last larval instar was found to reach a peak prior to that of the haemolymph ecdysteroid titre, supporting a role for the enzyme in development.

Responses of insect olfactory receptor neurons (ORNs) involve an entry of Ca2+ through olfactory heterodimeric receptor complexes. In moths, the termination of ORN responses was found to strongly depend on the external Ca2+ concentration through the activation of unknown Ca2+-dependent Cl− channels. We thus investigated the molecular identity of these Cl− channels. There is compelling evidence that bestrophins form Cl− channels when expressed in heterologous systems. Here we provide evidence that antennae of the moth Spodopteralittoralis express three transcripts encoding proteins with hallmarks of bestrophins. One of these transcripts, SlitBest1b, is expressed in ORNs. The heterologous expression of SlitBest1b protein in CHO-K1 cells yielded a Ca2+-activated Cl− current that shares electrophysiological properties with the native Ca2+-activated Cl− current of ORNs. Both currents are anionic, present similar dependence on the intracellular Ca2+ concentration, partly inactivate over time, have the same anion permeability sequence, the same sequence of inhibitory efficiency of blockers, the same almost linear I–V relationships and finally both currents do not depend on the cell volume. Therefore, our data suggest that SlitBest1b is a good candidate for being a molecular component of the olfactory Ca2+-activated Cl− channel and is likely to constitute part of the insect olfactory transduction pathway. A different function (e.g. regulation of other proteins, maintenance of the anionic homeostasis in the sensillar lymph) and a different role (e.g. involvement in the olfactory system development) cannot be excluded however. PMID:23300744

The male reproductive tract of Lepidoptera is an ideal model for the study of the physiological role of peripheral clocks in insects. The latter are significant in the generation and coordination of rhythmic phenomena which facilitate the initial stages of sperm capacitation. This process requires the maintenance of pH in the upper vas deferens (UVD) aided by, among others, H+-ATPase. Our aim was to determine the potential involvement of carbonic anhydrase (CA) in this process, an enzyme tasked with generating protons subsequently utilized by H+-ATPase to acidify the UVD milieu in S. littoralis, during the time when the lumen of this organ is filled with sperm. We attempted to answer the question whether CA activity can be controlled by the biological oscillator present in the male reproductive tract of the cotton leafworm. Using PAGE zymography, the presence of CA was demonstrated in the UVD wall, but not in the luminal fluid nor in the sperm. Using histochemistry, it was shown that CA is active in the UVD epithelium, and that this activity varies throughout the day and is most likely controlled by an endogenous biological clock. Conversely, the application of CA inhibitors, acetazolamide and sodium thiocyanate, in conjunction with an analysis of H+-ATPase activity in the acidification the UVD environment shows that CA most likely does not play a direct role in the regulation of the pH in this organ.

The naturally occurring phytoncidal chemical component of some plant- species are responsible for controlling and/or repelling insects from host plants. The use of natural products, readily available in the environment, to control the cotton leafworm, could be of help in reducing the need for applying the synthetic conventional insecticides . Moreover, crude vegetable oils application is quite safer to the non targets and the environment. The use of sex attractant pheromones as predicative tools to forecast pest population and their potential damage levels in specific crops, has significantly been limited because sex attractants are usually directed to one sex only. Thus, identification of plant constituents that attract S. littoralis Boisd. adults, especially females, and that directly affect their feeding or reproductive behaviour, would greatly expand opportunities for manipulation of the biological and environmental events that prevent the establishment, development, and dispersal of the insect-pest under study. Extracted Volatiles from certain parts of cotton plants, attracted both sexes of the cotton leafworm moths instead of attracting one sex only as the pheromones do. Plant flavonoids have been shown by many investigators to have an effect on insect behaviour, growth, and development. Quercetin is one of many bioflavonoids that exists in several fruits and vegetables. The Aim of the present work is to study in vivo the biochemical mode of action of quercetin as a synergist in combination with insecticides. The present results proved the synergistic effect of quercetin when combined with four insecticides, namely, profenofos (organophosphates), deltamethrin (pyrethroid), and tebufenozide et hexaflumuron as insect growth regulators against the studied insect-pest. Quercetin role as a synergist might be attributed to its ability to inhibit certain active oxidases, which may be responsible for in vivo detoxification of the intact insecticides when applied to

The third instars larvae of Spodotera littoralis were topically treated with two plant oils, Ricinus communis and Brassica nigra and one mineral oil, Kemesol 95% dissolved in petroleum ether and acetone at concentrations of 0.8, 1.6, 2.0, 3.0 & 4 %. The results revealed that the mean values of the total haemolymph and fat body protein was reduced in larvae treated with B. nigra and Kemesol 95%. A significant decrease was observed in haemolymph and fat body protein contents in larvae treated with all tested compound, the remarked decrease was noticed at the highest dose (4%) in both two solvents.

In an earlier study, we showed that the ingestive responses of the generalist caterpillar Spodopteralittoralis to foods imbalanced in their protein:carbohydrate content is similar to generalist locusts, but differs from that of specialist-feeding locusts. Here we further pursued the comparison by repeating the experiments using a closely related specialist caterpillar, Spodoptera exempta. First, caterpillars were allowed to self-compose a diet of preferred protein:carbohydrate balance by mixing between nutritionally complementary foods. Then, they were confined to one of five imbalanced foods, in which we measured the trade-off between over- and under-ingesting the two nutrients. On complementary foods, the caterpillars actively regulated their protein and carbohydrate intake. In the no-choice experiment, those fed excess-protein foods ingested small surpluses of protein compared with generalist feeders, thus showing a pattern of nutrient balancing similar to that observed in specialist locusts. Utilisation data indicated that ingested excesses and deficits were to some extent offset by differential utilisation. Evidence also showed that post-ingestive responses of the specialist S. exempta were less flexible than those observed in the generalist S. littoralis, a pattern which is again in accordance with comparisons of acridids differing in their host-plant range.

The fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), is an economically important pest of maize and other crops in the Americas. Studies suggest that modern varieties of maize lost some of their natural defense mechanisms against herbivores during domestication and agricultural selection. In the present study, we evaluated the attraction, feeding preference (host fidelity and consumption rate), and performance of S. frugiperda larvae reared on hybrid (Pioneer P4063W) and landrace (Tuxpeño) varieties of maize. We also evaluated the damage caused by S. frugiperda to Pioneer and Tuxpeño maize plants in the field. We found that fifth-instar larvae were more attracted to Pioneer plants than to Tuxpeño plants in a Y-tube olfactometer. Additionally, the fall armyworm larvae showed more fidelity to Pioneer leaves than to Tuxpeño leaves. However, the larval consumption rate was similar for both types of maize plants. The life cycle of S. frugiperda was significantly longer when the larvae were reared on Tuxpeño leaves than on Pioneer leaves. In the field, the Pioneer variety was infested with more S. frugiperda larvae than the Tuxpeño variety. Thus, our results provide evidence that modern varieties of maize may have lost some of their defensive traits during selective breeding.

Continuous mechanical damage initiates the rhythmic emission of volatiles in lima bean (Phaseolus lunatus) leaves; the emission resembles that induced by herbivore damage. The effect of diurnal versus nocturnal damage on the initiation of plant defense responses was investigated using MecWorm, a robotic device designed to reproduce tissue damage caused by herbivore attack. Lima bean leaves that were damaged by MecWorm during the photophase emitted maximal levels of β-ocimene and (Z)-3-hexenyl acetate in the late photophase. Leaves damaged during the dark phase responded with the nocturnal emission of (Z)-3-hexenyl acetate, but with only low amounts of β-ocimene; this emission was followed by an emission burst directly after the onset of light. In the presence of 13CO2, this light-dependent synthesis of β-ocimene resulted in incorporation of 75% to 85% of 13C, demonstrating that biosynthesis of β-ocimene is almost exclusively fueled by the photosynthetic fixation of CO2 along the plastidial 2-C-methyl-d-erythritol 4-P pathway. Jasmonic acid (JA) accumulated locally in direct response to the damage and led to immediate up-regulation of the P. lunatus β-ocimene synthase gene (PlOS) independent of the phase, that is, light or dark. Nocturnal damage caused significantly higher concentrations of JA (approximately 2–3 times) along with enhanced expression levels of PlOS. Transgenic Arabidopsis thaliana transformed with PlOS promoter∷β-glucuronidase fusion constructs confirmed expression of the enzyme at the wounded sites. In summary, damage-dependent JA levels directly control the expression level of PlOS, regardless of light or dark conditions, and photosynthesis is the major source for the early precursors of the 2-C-methyl-d-erythritol 4-P pathway. PMID:18165324

Dietary lipids, and in particular the essential fatty acids (EFA), EPA (20:5ω3) and DHA (22:6ω3), guarantee the well-being of animals and are recognized for their potential bottom-up control on animal populations. They are introduced in marine ecosystems through primary producers and when grazed upon, they are consumed, incorporated or modified by first-level consumers. As the availability of EFA in the ecosystem is affected by ambient temperature, the predicted rise in ocean temperature might alter the availability of these EFA at the basis of marine food webs. Despite the FA bioconversion capacity of certain benthic copepod species, their lipid (FA) response to varying temperatures is understudied. Therefore, the temperate, intertidal copepod Platychelipus littoralis was offered a mono and mixed diatom diet at 4, 15 °C (normal range) and at 24 °C (elevated temperature) to investigate the combined effects of temperature and resource availability on its FA content and composition. P. littoralis showed a flexible thermal acclimation response. Cold exposure increased the degree of FA unsaturation and the EPA%, and induced a shift towards shorter chain FA in the copepod's membranes. Furthermore, a mixed diet reduced the impact of heat stress on the copepod's membrane FA composition. Temperature affected the trophic transfer of EPA and DHA differently. While dietary resources could fully compensate for the temperature effects on total lipid and EPA content in the copepods, no such counterweigh was observed for the DHA dynamics. Heat stress lowered the DHA concentration in copepods regardless of the resources available and this implies negative effects for higher trophic levels.

Nitrogen (N) is one of the most critical chemical elements for plant and animal growth. Development and oviposition of the beet armyworm, Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae) was studied in relation to varying nitrogen levels in cotton, Gossypium hirsutum L. The development of S. exig...

Cyanogenic glucosides (CNglcs) are widespread plant defence compounds releasing toxic hydrogen cyanide when hydrolysed by specific β-glucosidases after plant tissue damage. In contrast to specialist herbivores that have mechanisms to avoid toxicity from CNglcs, it is generally assumed that non-adapted herbivores are negatively affected by CNglcs. Recent evidence, however, implies that the defence potential of CNglcs towards herbivores may not be as effective as previously anticipated. Here, performance, metabolism and excretion products of insects not adapted to CNglcs were analysed, including species with different degrees of dietary specialisation (generalists, specialists) and different feeding modes (leaf-snipping lepidopterans, piercing-sucking aphids). Insects were reared either on cyanogenic or acyanogenic plants or on an artificial cyanogenic diet. Lepidopteran generalists (Spodopteralittoralis, Spodoptera exigua, Mamestra brassicae) were compared to lepidopteran glucosinolate-specialists (Pieris rapae, Pieris brassicae, Plutella xylostella), and a generalist aphid (Myzus persicae) was compared to an aphid glucosinolate-specialist (Lipaphis erysimi). All insects were tolerant to cyanogenic plants; in lepidopterans tolerance was mainly due to excretion of intact CNglcs. The two Pieris species furthermore metabolized aromatic CNglcs to amino acid conjugates (Cys, Gly, Ser) and derivatives of these, which is similar to the metabolism of benzylglucosinolates in these species. Aphid species avoided uptake of CNglcs during feeding. Our results imply that non-adapted insects tolerate plant CNglcs either by keeping them intact for excretion, metabolizing them, or avoiding uptake.

Arabidopsis thaliana plants fend off insect attack by constitutive and inducible production of toxic metabolites, such as glucosinolates (GSs). A triple mutant lacking MYC2, MYC3, and MYC4, three basic helix-loop-helix transcription factors that are known to additively control jasmonate-related defense responses, was shown to have a highly reduced expression of GS biosynthesis genes. The myc2 myc3 myc4 (myc234) triple mutant was almost completely devoid of GS and was extremely susceptible to the generalist herbivore Spodopteralittoralis. On the contrary, the specialist Pieris brassicae was unaffected by the presence of GS and preferred to feed on wild-type plants. In addition, lack of GS in myc234 drastically modified S. littoralisfeeding behavior. Surprisingly, the expression of MYB factors known to regulate GS biosynthesis genes was not altered in myc234, suggesting that MYC2/MYC3/MYC4 are necessary for direct transcriptional activation of GS biosynthesis genes. To support this, chromatin immunoprecipitation analysis showed that MYC2 binds directly to the promoter of several GS biosynthesis genes in vivo. Furthermore, yeast two-hybrid and pull-down experiments indicated that MYC2/MYC3/MYC4 interact directly with GS-related MYBs. This specific MYC-MYB interaction plays a crucial role in the regulation of defense secondary metabolite production and underlines the importance of GS in shaping plant interactions with adapted and nonadapted herbivores.

In insects, like in other animals, experience-based modulation of preference, a form of phenotypic plasticity, is common in heterogeneous environments. However, the role of multiple fitness-relevant experiences on insect preference remains largely unexplored. For the multivoltine polyphagous moth Spodopteralittoralis we investigated effects of larval and adult experiences on subsequent reproductive behaviours. We demonstrate, for the first time in male and female insects, that mating experience on a plant modulates plant preference in subsequent reproductive behaviours, whereas exposure to the plant alone or plant together with sex pheromone does not affect this preference. When including larval feeding experiences, we found that both larval rearing and adult mating experiences modulate host plant preference. These findings represent the first evidence that host plant preferences in polyphagous insects are determined by a combination of innate preferences modulated by sensory feedback triggered by multiple rewarding experiences throughout their lifetime.

The beet armyworm, Spodoptera exigua (Hübner), has been anecdotally reported to oviposit more on drought stressed than on nonstressed cotton plants. Using potted cotton plants in cages, this study demonstrated that beet armyworms deposited 3.3, 4.6, and 2.3 times more (P < or = 0.05) eggs on cotton plants that were grown on 1500, 1000, and 750 ml water/wk, respectively, than on cotton plants grown in well watered (4000 ml water/wk) soil. Third instars, however, showed no preference for stressed cotton foliage over nonstressed foliage. Third instar beet armyworms raised on well watered cotton plants were 1.5, 2.3, and 2.6 times heavier than those reared on cotton grown in the 1500, 1000, and 750 ml water/wk plants (P < or = 0.05), respectively. Physiochemical analyses showed that drought stressed leaves had significantly greater accumulations of free amino acids that are essential for insect growth and development. Soluble protein and soluble carbohydrates were also more abundant in stressed leaves compared to nonstressed leaves. Despite the apparent increase in nutritional quality in drought stressed plants, larval survival was reduced, probably because the limiting factor became water. Greater amounts of cotton leaf area were consumed from drought stressed leaves (P < or = 0.05) than from nonstressed leaves, probably because the larvae had to metabolize greater portions of assimilated energy to supplement body water with metabolic water derived from respiration. The association of greater host plant nutritional quality to oviposition preference, and conversely, to reduced survivorship, is discussed.

Five alkaloids, in addition to hydroxytyrosol and uridine, were isolated from aerial parts of Alternanthera littoralis P. Beauv. Among the isolated compounds, alternamide A was an unusual tricyclic alkaloid with a bridged benzoazepine core. All isolated alkaloids have a catechol moiety, indicating a possible common biosynthetic route. Their structures were established by 1D and 2D NMR spectroscopy in combination with extensive tandem MS experiments by collisional induced dissociation (CID). The antiprotozoal activity of the isolated compounds was assayed against trypomastigote forms of Trypanosoma cruzi and amastigotes of Leishmania amazonensis. Alternamine A was the most active compound, reducing markedly the viability of both parasites. Antioxidant capacities evaluated by ORACFL assay showed that the isolated alkaloids (mainly alternamide B) contributed to the high activity recorded for the ethanolic crude extract; possibly, the catechol moiety present in all structures plays a central role in this result.

This study compared the cellular interactions of Spodopteralittoralis haemocytes with two virulence-different entomopathogenic fungi: Beauveriabassiana and Nomuraearileyi. Using light and transmission microscopy, five types of haemocytes namely, prohaemocytes (PRs), plasmatocytes (PLs), granulocytes (GRs), spherule cells (SPs) and oenocytoids (OEs) were identified in the 6th instar larvae. PRs and PLs were found in the haemopoietic tissue. Intra-haemocoelic injection of blastospores induced ultrastructural alterations in the cytoplasm and nuclei of circulating haemocytes of treated larvae. Different responses were observed in the populations of haemocyte types following injection with the tested fungi. The most important changes were the decrease of the numers of GRs accompanied with increase in SPs at 12-48h following injection with B. bassiana, whereas, a decrease of PLs with a commitment increase inSPs and OEs were observed at most time intervals after injection with N. rileyi. Both fungi provoked a decrease of the total number of haemocytes at 48h followed by an increase at 72h post-injection. In vivo assay showed that the GRs and PLs actively phagocytised fungal blastospores. There was a time-dependent decrease and increase in the phagocytosis activity after injection of B. bassiana and N. Rileyi, respectively. In B. bassiana-injected insects, the numbers nodules increased significantly at 6-48h in comparison with the controls post-injection. In N. rileyi-injected insects, nodules increased significantly only at 72h post-injection. No cellular encapsulation was observed in any of the examined insects.

A phytochemical investigation of the bulbs and flowers of Hymenocallis littoralis SALISB., cultivated in Egypt, was carried out, which resulted in the isolation of four alkaloids, lycorine (1), hippeastrine (2), 11-hydroxyvittatine (3), and (+)-8-O-demethylmaritidine (4), and of two flavonoids, quercetin 3'-O-glucoside (5), and rutin (6). The volatile constituents of the plant flowers were analyzed for the first time by GC/MS, which led to the identification of 26 known compounds (Table 1). Finally, the antimicrobial activity of the petroleum ether extract of the flowers of H. littoralis was investigated.

RNA interference is a potent and precise reverse genetic approach to carryout large-scale functional genomic studies in a given organism. During the past decade, RNAi has also emerged as an important investigative tool to understand the process of viral pathogenesis. Our laboratory has successfully generated transgenic reporter and RNAi sensor line of Spodoptera frugiperda (Sf21) cells and developed a reversal of silencing assay via siRNA or shRNA guided screening to investigate RNAi factors or viral pathogenic factors with extraordinary fidelity. Here we describe empirical approaches and conceptual understanding to execute successful RNAi screening in Spodoptera frugiperda 21-cell line.

Necrodes littoralis (Linnaeus, 1758) (Coleoptera: Silphidae), also known as the "shore sexton beetle," is a common silphid beetle that visits and breeds on large vertebrate cadavers. This study describes, for the first time, the involvement of N. littoralis on human corpses based on a large dataset of 154 French forensic cases. Various parameters regarding corpse location, decomposition stages, and entomofauna were extracted from each file. Compared to all of the forensic entomology cases analyzed between 1990 and 2013 (1028), N. littoralis was observed, on average, in one case out of eight; most of these cases occurred during spring and summer (73.5%). More than 90% of the cases were located outdoors, especially in woodlands, bushes, and fields. The decomposition stage of the corpse varied among cases, with more than 50% in the advanced decomposition stage, 36% in the early decomposition stage, and less than 10% in the fresh, mummified, or skeletonized stages. Regarding other necrophagous species sampled with N. littoralis, Calliphorid flies were found in 94% of the cases and Fanniidae/Muscidae in 65% of the cases. Chrysomya albiceps, a heliophilic species mostly located in the Mediterranean area, was present in 34% of the cases (only 20% in the whole dataset). The most common coleopteran species were Necrobia spp. (Coleoptera: Cleridae) and Creophilus maxillosus (Coleoptera: Staphylinidae); these beetles were observed in 27% of the cases. The over-representation of these species is likely due to similar requirements regarding the climate and decomposition stage. As N. littoralis is frequently observed and tends to become more common, we conclude that the developmental data for this species would be a precious tool for forensic entomologists in Europe.

The estimation of postmortem interval (PMI) based on successional patterns of adult insects is largely limited, due to the lack of potential PMI markers. Sex and size of adult insects could be easily used for such estimation. In this study, sex- and size-related patterns of carrion attendance by adult insects were analyzed in Necrodes littoralis (Coleoptera: Silphidae) and Creophilus maxillosus (Coleoptera: Staphylinidae). For both species, abundance of males and females changed similarly during decomposition. A slightly female-biased sex ratio was recorded in N. littoralis. Females of N. littoralis started visiting carcasses, on average, one day earlier than males. There was a rise in size of males of N. littoralis at the end of decomposition, whereas for females of both species and males of C. maxillosus, no size-related patterns of carrion visitation were found. Current results demonstrate that size and sex of adult carrion beetles are poor indicators of PMI.

By a orthogonal experiment, the influence of different ratio of phosphorus and potassium fertilizers on imperatorin, isoimperatorin and psoralen contents and yield of Glehnia littoralis were studied. The results showed that root dry weight and the yield of G. littoralis increased when reasonably applied phosphorus fertilizer combined with potassium fertilizer within a certain range. And the influence of phosphorus fertilizer was greater than that of potassium fertilizer. The optimal value of root dry weight and yield achieved at both P2O5 360 kg x hm(-2), K2O 270 kg x hm(-2) and P2O5 360 kg x hm(-2), K2O 180 kg x hm(-2). The effects of different phosphorus and potassium treatments on the content of imperatorin, isoimperatorin and psoralen in G. littoralis were determined, which shows that the content increased with the moderate increase of phosphorus and potassium. And the effects of phosphorus fertilizer were more significantly. The isoimperatorin content achieved the largest value at P2O5 360 kg x hm(-2), K2O 270 kg x hm(-2), also a larger content of imperatorin and psoralen. The imperatorin content is the largest when applied P2O5 360 kg x hm(-2), K2O 180 kg x hm(-2), and the isoimperatorin content was higher as well. So that the treatment of P2O5 360 kg x hm(-2), K2O 180 kg x hm(-2) are suitable for promote to the agricultural production, which could improve the quality and yield of G. littoralis.

Aloe littoralis Baker (Asphodelaceae family) is a well known plant in southern parts of Iran. Because of its use in Iranian folk medicine as a wound-healing agent, the present study was carried out to investigate anti-inflammatory and wound healing activities of this plant in Wistar rats. A. littoralis raw mucilaginous gel (ALRMG) and also two gel formulations prepared from the raw mucilaginous gel were used in this study. Gel formulations (12.5% and 100% v/w Aloe mucilage in a carbomer base) were applied topically (500 mg once daily) for 24 days in the thermal wound model. Also Aloe gel formulation (100%) and ALRMG (500 mg daily) were evaluated in incisional wound model. Carrageenan-induced paw edema was used to assess the anti-inflammatory effect of intraperitoneal injection of ALRMG. In burn wound, ALRMG and Aloe formulated gel (100%) showed significant (P<0.05) healing effect. Topical application of ALMRG and Aloe formulated gel (100%) promoted healing rate of incisional wound. In carrageenan test, ALRMG (2.5 and 5 ml/Kg) revealed significant (P<0.05) anti-inflammatory activity. Results showed that A. littoralis is a potential wound-healing and anti-inflammatory agent in rats. Further studies are needed to find out the mechanism of these biological effects and also the active constituents responsible for the effects.

We studied germination behaviors and persistence mechanism of wild Glehnia littoralis, a typical coastal species at temperate sandy coasts of the North Pacific Ocean, and tested the hypothesis that the coastal plants may have evolved special seeds adapting to the coasts, by which they recruit and persist easily, occupying the coasts as ideal habitats. In the Shandong Peninsula, China, we investigated temperature and moisture conditions of coast sand in relation to germination and evaluated effects of sand burial, seawater immersion and sowing time on germination. When germination began, daily dawn temperatures of sand were about 10 °C and daily noon temperatures were about 25 °C; the temperatures were not different in the sand <8 cm deep. The sand at these depths showed a significant difference in moisture contents. The seeds exhibited large germination rates if sand burial was at depths >= 3 cm and winter freezing was kept longer than 2.5 months. Seeds experiencing seawater immersion were able to germinate well. These evidences suggest that G. littoralis has evolved special seeds adapting to seawater dispersal and specific season rhythm. By the seeds, G. littoralis occupies temperate sandy coasts as ideal habitats to persist. PMID:28211487

Vegetative insecticidal proteins (Vips) are insecticidal proteins synthesized by Bacillus thuringiensis during the vegetative stage of growth. In this study, Vip3Aa protein, obtained by in vitro expression of the vip3Aa gene from B. thuringiensis WB5, displayed high insecticidal activity against Spodoptera litura aside from Spodoptera exigua and Helicoverpa armigera. Bioassay results showed that the toxicity of Vip3Aa protein against S. litura larvae statistically decreased along with the increase of the age of the larvae, with LC50 = 2.609 ng/cm(2) for neonatal larvae, LC50 = 28.778 ng/cm(2) for first instar larvae, LC50 = 70.460 ng/cm(2) for second instar larvae, and LC50 = 200.627 ng/cm(2) for third instar larvae. The accumulative mortality of 100% larvae appeared at 72 h for all instars of S. litura larvae, when feeding respectively with 83.22, 213.04, 341.40, and 613.20 ng/cm(2) of Vip3Aa toxin to the neonatal and first to third instar larvae. The histopathological effects of Vip3Aa toxin on the midgut epithelial cells of S. litura larvae was also investigated. The TEM observations showed wide damage of the epithelial cell in the midgut of S. litura larvae fed with Vip3Aa toxin.

A recombinant baculovirus, rv-egfp-NK, containing a reporter gene encoding the enhanced green fluorescent protein (EGFP), was used to express nattokinase (NK), a fibrinolytic enzyme, in Spodoptera frugiperda (SF-9) cells. The recombinant protein also included a histidine tag for purification using Ni(2+) resins. The recombinant NK, approximately 30 kDa, retained fibrinolytic activity (60 U/ml). The integration of the EGFP expression cassette in the Bac-to-Bac system is thus an effective method for the expression and purification of recombinant NK protein in Spodoptera frugiperda insect cells.

Background: Linear dermatitis is endemic in Iran where most cases occur in the Caspian Sea coast and Fars province. The disease is caused by beetles of the genus Paederus which are active from early spring to beginning of autumn although its incidence rises from May to August. The classic taxonomy of Paederus spp. is based on the male genitalia that is very complex and needs expertise. In this study, we report a DNA-based method to discriminate Paederus fuscipes and Paederus littoralis (=syn: P. lenkoranus, P. ilsae). Methods: Type specimens were collected from north and south of Iran. Molecular typing of the species was performed using restriction fragment length polymorphism (RFLP) analysis of polymerase chain reaction (PCR)-amplified fragments of mtDNA-COI. Results: Sequence analyses of the data obtained in this study showed significant DNA polymorphisms. There were 89 substitutions between COI sequences of the two species. The mtDNA-COI fragment comprises several useful species-specific restriction sites comprising HaeIII that could result in distinctively different species-specific PCR–RFLP profiles. The HaeIII enzyme cuts the 872 bp PCR amplicon of P. littoralis into 737 and 100 bp and two small nonvisible bands whereas it does not cut P. fuscipes amplicon into fragments. Conclusion: This study demonstrates that molecular typing is useful method and allows one to differentiate between two species and is recommended for discrimination of other Paederus species, which morphologically are indistinguishable or very difficult to be distinguished. PMID:28032097

Abstract- Spodoptera frugiperda consists of two genetically and behaviorally different strains, the corn- and the rice-strain, which seem to be in the process of sympatric speciation. We investigated the role of strain-specific sexual communication as a prezygotic mating barrier between both strains...

Shortly prior to death, many species of Lepidoptera infected with nucleopolyhedrovirus climb upwards on the host plant. This results in improved dissemination of viral occlusion bodies over plant foliage and an increased probability of transmission to healthy conspecific larvae. Following applications of Spodoptera exigua multiple nucleopolyhedrovirus for control of Spodoptera exigua on greenhouse-grown sweet pepper crops, necrophagy was observed by healthy S. exigua larvae that fed on virus-killed conspecifics. We examined whether this risky behavior was induced by olfactory or phagostimulant compounds associated with infected cadavers. Laboratory choice tests and olfactometer studies, involving infected and non-infected cadavers placed on spinach leaf discs, revealed no evidence for greater attraction of healthy larvae to virus-killed over non-infected cadavers. Physical contact or feeding on infected cadavers resulted in a very high incidence of transmission (82–93% lethal disease). Observations on the behavior of S. exigua larvae on pepper plants revealed that infected insects died on the uppermost 10% of foliage and closer to the plant stem than healthy conspecifics of the same stage, which we considered clear evidence of baculovirus-induced climbing behavior. Healthy larvae that subsequently foraged on the plant were more frequently observed closer to the infected than the non-infected cadaver. Healthy larvae also encountered and fed on infected cadavers significantly more frequently and more rapidly than larvae that fed on non-infected cadavers. Intraspecific necrophagy on infected cadavers invariably resulted in virus transmission and death of the necrophagous insect. We conclude that, in addition to improving the dissemination of virus particles over plant foliage, baculovirus-induced climbing behavior increases the incidence of intraspecific necrophagy in S. exigua, which is the most efficient mechanism of transmission of this lethal pathogen. PMID

The essential oils from the cladodes of Opuntia littoralis, Opuntia ficus-indica and Opuntia prolifera growing wild on Santa Catalina Island, California, were obtained by hydrodistillation and analysed by gas chromatography-mass spectrometry (GC-MS). Terpenoids were the dominant class of volatiles in O. littoralis, with the two main components being the furanoid forms of cis-linalool oxide (10.8%) and trans-linalool oxide (8.8%). Fatty acid-derived compounds dominated the essential oil of O. ficus-indica with linoleic acid (22.3%), palmitic acid (12.7%), lauric acid (10.5%) and myristic acid (4.2%) as major fatty acids. O. prolifera oil was composed of 46.6% alkanes and the primary hydrocarbon component was heptadecane (19.2%). Sixteen compounds were common to all the three Opuntia species.

TEXAS Galveston Bay GU LF OF ME XI CO In cooperation with the U.S. Fish and Wildlife Service Occurrence of the Diamondback Terrapin (Malaclemys...Occurrence of the Diamondback Terrapin (Malaclemys Terrapin Littoralis) at South Deer Island in Galveston Bay, Texas, April 2001-May 2002 5a. CONTRACT...of Diamondback terrapin by L.S. Coplin, U.S. Geological Survey. U.S. Department of the Interior U.S. Geological Survey Occurrence of the Diamondback

Particle-induced X-ray emission and particle-induced gamma-ray emission spectrometry were successfully applied in a study of the elemental composition of decomposing filamentous algae. Fresh brown (Pilayella littoralis) and green (Cladophora glomerata) algal materials were placed in cages at 4m depth in a water column of 8m in the Archipelago Sea, northern Baltic Sea. Every second week decaying algae were sampled from the cages to allow measurements of changes in the elemental compositions. In the study of the elemental losses the concentrations were compensated for the mass reduction. The results show that sulphur, chlorine and partly potassium were lost during decomposition of P. littoralis and C. glomerata. Most of the other elements studied were recovered in the remaining algal mass. Special attention was paid to sorption and desorption of elements, including metal binding capacity, in the decaying algal materials. The affinity order of different cations to the two algal species was established by calculation of conditional distribution coefficients, D'(M). For instance for P. littoralis the following series of binding strength (affinity) of cations were obtained: Al>Ti>Fe > Mn>Ni, Cu>Ba, Cr, Zn>Rb>K, Sr>Pb>Ca>Na>Mg. Notably is that the binding strength of strontium was more than 10 times higher for P. littoralis than for C. glomerata. Due to their high binding capacity and good affinity and selectivity for heavy metal ions these algae have great potential as biological sorbents. Large variations in elemental content during decomposition complicate the use of algae for environmental monitoring.

In this study, crude lantadene was extracted from Lantana canmara leaves, and its antifeeding effects on Plutella xylostella and Spodoptera litura larvae were tested. In no-choice test, crude lantadene at 1.6 mg x ml(-1) concentration had antifeeding effects on the 2nd instar P. xylostella larvae and 1st instar S. litura larvae, with the antifeeding rate being 62.4% and 33.1%, respectively within 48 h. In choice test, even a low concentration (0.4 mg x ml(-1)) crude lantadene still had anti-feeding effects on the 2nd instar P. xylostella larvae, and the antifeeding rate at 0.4, 0.8 and 1.6 mg x ml(-1) concentration was 52.7%, 55.5% and 78.9%, respectively. Crude lantadene only at 1.6 mg x ml(-1) concentration had anti-feeding effects on the 1st instar S. litura larvae, and the antifeeding rate was 33.0%. For the 2nd instar S. litura larvae, crude lantadene had no antifeeding effects both in no-choice and in choice test.

Citrus aurantium and C. limon were selected in the search for natural plant insecticides. The essential oils of C. aurantium and C. limon and ethanol extracts of the seeds, pulp, albedo, and peel of C. aurantium were incorporated into the larval diet of the lepidopteran pest Spodoptera frugiperda. Larval and pupal mortality were quantified and adult malformation was observed. C aurantium essential oil had antifeedant action and the mixture of albedo ethanol extract and C aurantium essential oil had toxic effects on S. frugiperda larvae at early stages, when they had not yet produced major damage to the crop. Our results indicated that a mixture of ethanol extract of albedo and C. aurantium essential oil (250 microg of extract mix per g of diet) deterred feeding by 46% and had the highest larval mortality (100%) of the materials tested. The peel extract (250 microg per g of diet) produced an increment in growth rate and diet consumption. However, 40% of the larval and 45% of the pupal populations died after 96 h of treatment. The blend of essential oil and C. aurantium albedo ethanol extract showed the lowest consumption and a poor nutrient conversion into biomass. Finally, the presence of D-limonene and nootkatone in the peel ethanol extract, and C. limon and C. aurantium essential oils, may be the cause of the response in the feeding behavior and toxic effects found on S. frugiperda.

The fall armyworm [Spodoptera frugiperda (J.E. Smith)] is a serious pest of many field and horticulture crops. Because of the many advantages for the use of plant-derived pesticides, we tested whether bee balm (Monarda didyma L.) leaves could have feeding deterrence on fall armyworm. When S. frugipe...

The feeding of Spodopteralittoralis and Autographa gamma caterpillars on tansy leaves led to a complete different release of volatile monoterpenes, sesquiterpenes, and hexenyl alkanoates. Volatiles were collected from S. littoralis and A. gamma larvae damaged, mechanically wounded, and excised tansy leaves by closed loop stripping analysis. The qualitative and quantitative determination of the volatiles were done by GC-MS- and GC-measurements. The oligosaccharides sucrose, raffinose, stachyose, and verbascose have been detected in oral secretion of the caterpillars of the cotton leafworm S. littoralis. When applied to damaged leaves of tansy plants, these oligosaccharides induce the tansy leaves to emit a similar volatile blend as the feeding of S. littoralis larvae.

The Hymenocallis littoralis, an ornamental and medicinal plant, had been traditionally used for wound healing. In the present study, an analytical method using HPLC with ultraviolet detection was developed for the quantification of lycorine in the extracts of different parts of wild plant and tissue culture samples of H. littoralis. The separation was achieved using a reversed-phase column. The method was found to be accurate, repeatable, and sensitive for the quantification of minute amount of lycorine present in the samples. The highest lycorine content was found in the bulb extract (2.54 ± 0.02 μg/mg) whereas the least was in the root extract (0.71 ± 0.02 μg/mg) of the wild plants. Few callus culture samples had high content of lycorine, comparable to that of wild plants. The results showed that plant growth regulators, 2,4-dichlorophenoxyacetic acid (2,4-D) alone at 4.5 μM (2.58 ± 0.38 μg/mg) or a combination of 2,4-D at 9.00 μM with 4.5 μM of 6-benzylaminopurine (BAP), were the optimum concentrations for the production of high lycorine (2.45 ± 0.15 μg/mg) content in callus culture. The present analytical method could be of value for routine quantification of lycorine in the tissue culture production and standardization of the raw material or extracts of H. littoralis.

The host selection for oviposition by Spodoptera frugiperda (J.E. Smith) among corn, millet, cotton and soybean, and its relationship with the biological characteristics were investigated. Free and non-choice tests for oviposition using plots containing five plants each, from each host in plastic greenhouse, resulted in similar oviposition preference among the host plants. In addition, selected biological characteristics of S. frugiperda were determined in the laboratory with larvae feeding on host leaves, and the combination of leaf and cotton boll. Neonate larvae exhibited low success of colonization on cotton boll compared to the leaves of all other hosts. Spodoptera frugiperda fed only on cotton bolls exhibited longer larval and pupal development, and longer adult life span; however with similar egg production. Larvae fed cotton leaves during six days and then transferred to cotton bolls, however, exhibited development and reproduction similar to those reared on corn or only on cotton leaves. Therefore, the variations on immature stages of S. frugiperda were not related with host selection for oviposition which was similar among the studied hosts. Based on our data, the millet as a winter, rotational, and cover crop is a potential host for S. frugiperda, while leaves and cotton bolls were diets of intermediate suitability as compared to corn and soybean leaves.

Cannibalism in the fall armyworm, Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) (FAW), is a limiting factor in a baculovirus production system. To detect the impact of cannibalism, a two-step bioassay was conducted with different larval ages of FAW fed on two food sources (corn and castor bean leaves) contaminated with the S. frugiperda multiple-embedded nucleopolyhedrovirus. In a first bioassay, the food source affected the cannibalism, being higher for all larval ages tested (5-, 6- and 7-day-old larvae) in larvae fed on corn than on those fed on castor bean leaves. Larval mortality, weight equivalent and larval equivalents (LEs) per hectare decreased as the larval age increased. Larval weight, occlusion bodies (OBs)/larva and total OBs increased when the larval age increased. In a second bioassay, in which only 6- and 7-day-old larvae were used because of the performance in the first bioassay, the cannibalism rates were affected by the interaction between food sources and time of feeding (48 and 72 h), reaching the highest values for 6- and 7-day-old larvae fed on corn leaves for 72 h. Mortality of the FAW was affected by the interaction between food sources, larval age and time of feeding. The lowest mortalities were on 7-day-old larvae when they were fed on castor bean leaves for 48 and 72 h. Larval weight, OBs/larva, total OBs and LEs were affected by the interaction between food sources and larval age. A significant correlation was observed between larval weight and OBs/larva that fed on both food sources, suggesting that larval weight can be used to achieve a concentration to be sprayed in 1 ha.

Hexane and ethanol extracts of seeds from 22 species of plants of the family Meliaceae from a number of countries were prepared. The extracts were submitted to antifeedant and toxicity bioassays utilizing fall armyworm [Spodoptera frugiperda (J.E. Smith)] (Lepidoptera: Noctuidae) larvae and striped cucumber beetle [Acalymma vittatum (F.)] (Coleoptera: Chrysomelidae) adults. Toxicity tests were also performed with brine shrimp,Anemia salina Leach. Feeding inhibition and mortality produced by some of these extracts were comparable to and, in certain cases, slightly greater than the effects produced by comparable neem (Azadiracta indica A. Juss.) seed preparations. Brine shrimp toxicity data do not extrapolate to insect activity, and vice versa.

The repulsive activity of Cestrum parquii on the feeding behaviour of Pieris brassicae L. and Spodopteralittoralis Boisduval was studied. Antifeedant effect was observed at regular intervals during a week. Meal uptake was stopped after the first day, showing a strongly antifeedant effect of C. parquii extract on these two species. The impact of C. parquii with different concentrations of dry extract (2, 4, 8, 16, 32%) delayed the larval growth of Spodopteralittoralis Boisduval compared to untreated insects. Abnormal metamorphosis was observed with concentrations of 8 and 16% leading to death and abnormal morphogenesis. The antifeedant activity of C. parquii was also observed in S. littoralis and measured by the evolution of food uptake and the weight of larva. Effect seemed to be proportional to extract concentration in the diet. High concentration (32%) stopped growth in the earlier instar.

Plants exhibit multifarious defence traits against herbivory that are constitutively expressed or induced upon attack. Insect egg deposition often precedes impending larval attack, and several plants can increase their resistance against larvae after experiencing the oviposition by an herbivore. The nature of such oviposition-mediated resistance remains unknown, and here we aim to determine plant traits that explain it. We test whether oviposition on a host plant can induce plant defence responses or enhance (prime) the induction of defence traits in response to larval herbivory. We exposed Nicotiana attenuata plants to oviposition by moths of a generalist herbivore, Spodoptera exigua. Its larvae suffered higher mortality, retarded development and inflicted less feeding damage on oviposition-experienced than on oviposition-unexperienced plants. While oviposition alone did not induce any of the examined defence traits, oviposited plants exhibited a stronger inducibility of known defence traits, i.e. caffeoylputrescine (CP) and trypsin protease inhibitors (TPIs). We found no effects of oviposition on phytohormone levels, but on the feeding-inducible accumulation of the transcription factor NaMyb8 that is governing biosynthesis of phenylpropanoid-polyamine conjugates, including CP. Comparison of larval performance on wild-type plants, CP-deficient plants (silenced NaMyb8 gene), and TPI-deficient plants (silenced NaPI gene) revealed that priming of plant resistance to larvae by prior oviposition required NaMyb8-mediated defence traits. Our results show that plants can use insect egg deposition as a warning signal to prime their feeding-induced defence.

We describe the nest, eggs, and nestlings of the Restinga Antwren (Formicivora littoralis), an endangered bird of Restinga ecosystem (sandy coastal plain vegetation) that is endemic to Rio de Janeiro state. Twelve nests were found at the edges of trails or natural gaps at Massambaba Restinga region, in different supporting plants and heights from the ground (X ± SD 1.27 ± 0.97 m, range 0.27 to 3.45 m). Nests were cup-shaped and were in horizontal forks attached to branches at three to five points with whitish, soft, and thin cotton-like vegetable fiber. The nests' cup shape and measurements were similar to congeneric species, but nest material was different. Eggs were white with brown spots concentrated on the large end or around the middle, giving the appearance of a rough brown ring. Their mean (± SD) minimum diameter was 13.1 ± 0.34 mm, with maximum diameter of 18.0 ± 0.38 mm, and mass of 1.7 ± 0.18 g (n = 8). We found two nestlings completely naked on their first day after hatching.

Island endemics are typically differentiated from their mainland progenitors in behavior, morphology, and genetics, often resulting from long-term evolutionary change. To examine mechanisms for the origins of island endemism, we present a phylogeographic analysis of whole mitochondrial genomes from the endangered island fox (Urocyon littoralis), endemic to California’s Channel Islands, and mainland gray foxes (U. cinereoargenteus). Previous genetic studies suggested that foxes first appeared on the islands >16,000 years ago, before human arrival (~13,000 cal BP), while archaeological and paleontological data supported a colonization >7000 cal BP. Our results are consistent with initial fox colonization of the northern islands probably by rafting or human introduction ~9200–7100 years ago, followed quickly by human translocation of foxes from the northern to southern Channel Islands. Mitogenomes indicate that island foxes are monophyletic and most closely related to gray foxes from northern California that likely experienced a Holocene climate-induced range shift. Our data document rapid morphological evolution of island foxes (in ~2000 years or less). Despite evidence for bottlenecks, island foxes have generated and maintained multiple mitochondrial haplotypes. This study highlights the intertwined evolutionary history of island foxes and humans, and illustrates a new approach for investigating the evolutionary histories of other island endemics. PMID:25714775

The island fox ( Urocyon littoralis) is one of few reportedly endemic terrestrial mammals on California's Channel Islands. Questions remain about how and when foxes first colonized the islands, with researchers speculating on a natural, human-assisted, or combined dispersal during the late Pleistocene and/or Holocene. A natural dispersal of foxes to the northern Channel Islands has been supported by reports of a few fox bones from late Pleistocene paleontological localities. Direct AMS 14C dating of these "fossil" fox bones produced dates ranging from ˜ 6400 to 200 cal yr BP, however, postdating human colonization of the islands by several millennia. Although one of these specimens is the earliest securely dated fox from the islands, these new data support the hypothesis that Native Americans introduced foxes to all the Channel Islands in the early to middle Holocene. However, a natural dispersal for the original island colonization cannot be ruled out until further paleontological, archaeological, and genetic studies (especially aDNA [ancient DNA]) are conducted.

Island endemics are typically differentiated from their mainland progenitors in behavior, morphology, and genetics, often resulting from long-term evolutionary change. To examine mechanisms for the origins of island endemism, we present a phylogeographic analysis of whole mitochondrial genomes from the endangered island fox (Urocyon littoralis), endemic to California's Channel Islands, and mainland gray foxes (U. cinereoargenteus). Previous genetic studies suggested that foxes first appeared on the islands >16,000 years ago, before human arrival (~13,000 cal BP), while archaeological and paleontological data supported a colonization >7000 cal BP. Our results are consistent with initial fox colonization of the northern islands probably by rafting or human introduction ~9200-7100 years ago, followed quickly by human translocation of foxes from the northern to southern Channel Islands. Mitogenomes indicate that island foxes are monophyletic and most closely related to gray foxes from northern California that likely experienced a Holocene climate-induced range shift. Our data document rapid morphological evolution of island foxes (in ~2000 years or less). Despite evidence for bottlenecks, island foxes have generated and maintained multiple mitochondrial haplotypes. This study highlights the intertwined evolutionary history of island foxes and humans, and illustrates a new approach for investigating the evolutionary histories of other island endemics.

In this study , two strains of Bacillus belonging to two serotypes and four of their transconjugants were screened with respect to their toxicity against lepidopterous cotton pest. . Bacterial transconjugants isolated from conjugation between both strains were evaluated for their transconjugant efficiency caused mortality in Spodopteralittoralis larvae . Two groups of bioinsecticides ; crystals , crystals and spores have been isolated from Bacillusstrains and their transconjugants . Insecticidal crystal protein ( ICP ) was specific for lepidopteran insects because of the toxin sufficient both for insect specificity and toxicity . The toxicities of these two groups against larvae of Spodopteralittoralis was expressed as transconjugant efficiency , which related to the mean number of larvae died expressed as mortality percentage . The results showed transconjugant efficiency in reducing the mean number of Spodopteralittoralis larvae feeding on leaves of Ricinus communis sprayed with bioinsecticides of Bt transconjugants. Most values of positive transconjugant efficiency related to increasing mortality percentage are due to toxicological effects appeared in response to the treatments with crystals + endospores than that of crystals alone .This indicated that crystals + endospores was more effective for increasing mortality percentage than that resulted by crystals . Higher positive transconjugant efficiency in relation to the mid parents and better parent was appeared at 168 h of treatment . The results indicated that recombinant Bacillus thuringiensis are important control agents for lepidopteran pests , as well as , susceptibility decreased with larval development . The results also suggested a potential for the deployment of these recominant entomopathogens in the management of Spodoptera. littoralis larvae .

Proteinase inhibitors (PIs) from the seeds of bitter gourd (Momordica charantia L.) were identified as strong inhibitors of Helicoverpa armigera gut proteinases (HGP). Biochemical investigations showed that bitter gourd PIs (BGPIs) inhibited more than 80% HGP activity. Electrophoretic analysis revealed the presence of two major proteins (BGPI-1 and-2) and two minor proteins (BGPI-3 and-4) having inhibitory activity against both trypsin and HGP. The major isoforms BGPI-1 and BGPI-2 have molecular mass of 3.5 and 3.0 kDa, respectively. BGPIs inhibited HGP activity of larvae fed on different host plants, on artificial diet with or without added PIs and proteinases excreted in fecal matter. Degradation of BGPI-1 by HGP showed direct correlation with accumulation of BGPI-2-like peptide, which remained stable and active against high concentrations of HGP up to 3 h. Chemical inhibitors of serine proteinases offered partial protection to BGPI-1 from degradation by HGP, suggesting that trypsin and chymotrypsin like proteinases are involved in degradation of BGPI-1. In larval feeding studies, BGPIs were found to retard growth and development of two lepidopteran pests namely Helicoverpa armigera and Spodoptera litura. This is the first report showing that BGPIs mediated inhibition of insect gut proteinases directly affects fertility and fecundity of both H. armigera and S. litura. The results advocate use of BGPIs to introduce insect resistance in otherwise susceptible plants.

Fungal endosymbionts in plants may influence interactions among plants, herbivores and their parasitoids through the production of secondary metabolites. We used a lepidopteran pest and its generalist parasitoid to test the effect of endophyte-infected plants on a third trophic level. Endophytic fungi, Aspergillus flavus and Aspergillus niger, isolated from Acacia arabica, were used to infect cauliflower plants. We found that the presence of the endophyte in the plants significantly extended the development period of Spodoptera litura (Fab.) larvae. Feeding of the host on endophyte-infected plants further adversely affected the development and performance of its parasitoid, Bracon hebetor (Say). A negative impact was also recorded for longevity and fecundity of endophyte-naive parasitoid females due to the parasitization of host larvae fed on endophyte-infected plants. The presence of endophytes in the diet of the host larvae significantly prolonged the development of the parasitoid. A strong detrimental effect was also recorded for larval survival and emergence of parasitoid adults. The longevity and parasitism rate of female wasps were reduced significantly due to the ingestion of endophyte-infected cauliflower plants by S. litura larvae. Overall, we found that both endophytic fungi had a negative impact on the parasitoid.

Spodoptera exigua, which feeds on various crops worldwide, has natural enemies that are susceptible to the insecticides used against S. exigua. We investigate the toxicity and residue risk of 9 insecticides on the development of H. axyridis, C. sinica, S. manilae and T. remus. S. manilae and T. remus adults were sensitive to all 9 insecticides (LC50 less than 2.75 mg a.i. liter(-1)), while H. axyridis and C. sinica adults were less sensitive (LC50 between 6 × 10(-5) mg a.i. liter(-1) and 78.95 mg a.i. liter(-1)). Emamectin benzoate, spinosad, indoxacarb, alpha-cypermethrin, chlorfenapyr and chlorantraniliprole showed no toxicity on H. axyridis, C. sinica, S. manilae and T. remus pupae with the recommended field concentrations. The risk analysis indicated that chlorantraniliprole is harmless to larvae of four natural enemies and adult of H. axyridis, C. sinica and S. manilae. Emamectin benzoate and spinosad had higher safety to the development of H. axyridis, C. sinica, S. manilae and T. remus with the risk duration less than 4d. Indoxacarb, tebufenozide, chlorfenapyr, methomyl, alpha-cypermethrin and chlorpyrifos showed dangerously toxic and long risk duration on S. manilae and T. remus adults.

Spodoptera exigua, which feeds on various crops worldwide, has natural enemies that are susceptible to the insecticides used against S. exigua. We investigate the toxicity and residue risk of 9 insecticides on the development of H. axyridis, C. sinica, S. manilae and T. remus. S. manilae and T. remus adults were sensitive to all 9 insecticides (LC50 less than 2.75 mg a.i. liter−1), while H. axyridis and C. sinica adults were less sensitive (LC50 between 6 × 10−5 mg a.i. liter−1 and 78.95 mg a.i. liter−1). Emamectin benzoate, spinosad, indoxacarb, alpha-cypermethrin, chlorfenapyr and chlorantraniliprole showed no toxicity on H. axyridis, C. sinica, S. manilae and T. remus pupae with the recommended field concentrations. The risk analysis indicated that chlorantraniliprole is harmless to larvae of four natural enemies and adult of H. axyridis, C. sinica and S. manilae. Emamectin benzoate and spinosad had higher safety to the development of H. axyridis, C. sinica, S. manilae and T. remus with the risk duration less than 4d. Indoxacarb, tebufenozide, chlorfenapyr, methomyl, alpha-cypermethrin and chlorpyrifos showed dangerously toxic and long risk duration on S. manilae and T. remus adults. PMID:27958333

Melanism has been found in a wide range of species, but the molecular mechanisms involved remain largely elusive. In this study, we studied the molecular mechanisms of the pupal melanism in Spodoptera exigua. The full length cDNA sequences of tyrosine hydroxylase (TH) and dopa decarboxylase (DDC), two key enzymes in the biosynthesis pathway of melanin, were cloned, and their temporal expression patterns in the integument were compared during the larval-pupal metamorphosis process of the S. exigua wild type (SEW) and melanic mutant (SEM) strains. No amino acid change in the protein sequence of TH and DDC was found between the two strains. Both DDC and TH were significantly over-expressed in the integument of the SEM strain at late-prepupa and 0 h pupa, respectively, compared with those of the SEW strain. Feeding 5th instar larvae of SEM with diets incorporated with 1 mg/g of the DDC inhibitor L-α-Methyl-DOPA and 0.75 mg/g of the TH inhibitor 3-iodo-tyrosine (3-IT) resulted in 20% pupae with partially-rescued phenotype and 68.2% of pupae with partially- or fully-rescued phenotype, respectively. These results indicate that overexpressions of TH and DDC are involved in the pupal melanization of S. exigua. PMID:26084938

The wild soybean (Glycine soja Sieb. et Zucc) has been reported to be relatively resistant to insect and pathogenic pests. However, the responsible secondary metabolites in the aerial part of this important plant are largely unknown. From the aerial part of G. soja, 13 compounds were isolated and identified, including seven isoflavonoids (1-7), a cyclitol (8), two sterol derivatives (9 and 10), and three triterpenoids (11-13). Compound 7 is a new isoflavonoid, and compounds 9 and 10 are reported as natural products for the first time. The growth inhibitory activity of 1, 3, 4, and 8 against the larvae of Spodoptera litura was investigated. The most abundant isoflavonoid in the aerial part of G. soja, daidzein (1), which could not be metabolized by S. litura, was found to inhibit the insect larvae growth significantly in 3 days after feeding diets containing the compound. Compounds 3, 4, and 8, which could be partially or completely metabolized, were inactive. Our results suggested that the isoflavonoid daidzein (1) might function as a constitutive defense component in G. soja against insect pests.

The fall armyworm, Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) is an economically significant pest of sorghum and maize. To screen sorghum and maize germplasm for resistance to fall armyworm feeding, field, greenhouse, or lab bioassays are often utilized individually or in combinatio...

The Hymenocallis littoralis, an ornamental and medicinal plant, had been traditionally used for wound healing. In the present study, an analytical method using HPLC with ultraviolet detection was developed for the quantification of lycorine in the extracts of different parts of wild plant and tissue culture samples of H. littoralis. The separation was achieved using a reversed-phase column. The method was found to be accurate, repeatable, and sensitive for the quantification of minute amount of lycorine present in the samples. The highest lycorine content was found in the bulb extract (2.54 ± 0.02 μg/mg) whereas the least was in the root extract (0.71 ± 0.02 μg/mg) of the wild plants. Few callus culture samples had high content of lycorine, comparable to that of wild plants. The results showed that plant growth regulators, 2,4-dichlorophenoxyacetic acid (2,4-D) alone at 4.5 μM (2.58 ± 0.38 μg/mg) or a combination of 2,4-D at 9.00 μM with 4.5 μM of 6-benzylaminopurine (BAP), were the optimum concentrations for the production of high lycorine (2.45 ± 0.15 μg/mg) content in callus culture. The present analytical method could be of value for routine quantification of lycorine in the tissue culture production and standardization of the raw material or extracts of H. littoralis. PMID:24895650

Hymenocallis littoralis growing in southern China has been recently extensively damaged by virus-like symptoms of necrosis, chlorosis, and ringspot. Of 44 plant samples collected from Yunnan, Guangxi, Guangdong, and Fujian provinces in southern China, 32 were infected with hippeastrum chlorotic ringspot virus (HCRV). Phylogenetic analysis based on the N gene divided the virus samples into two branches suggesting a geographic distribution attributed to the initial stage of a founder effect. The N gene was under purifying selection pressure and most of the deleterious mutants had been removed. Both the population dynamics and genetic analyses suggested that populations of HCRV in southern China are spreading.

Advances in techniques for rearing insects on artificial diets are fundamental to solving issues of basic and applied entomology. In this study, we evaluated the development of Spodoptera albula (Walker) (Lepidoptera: Noctuidae) on three artificial diets used for other species of Lepidoptera, at three larval densities, and two densities of adult couples housed in oviposition cages of two sizes, with the aim of optimizing methodology for rearing S. albula in the laboratory. Biological parameters were recorded from S. albula, and a fitness index was calculated based on the larval survival and duration and weight of pupae. The total and daily oviposition was recorded using 5 or 10 adult couples of S. albula housed in two cage sizes. Concentrations of total nitrogen and protein in the tested diets were determined. Development of S. albula was completed in all artificial diets; however, the diet used for rearing Anticarsia gemmatalis (Hübner) larvae was the most suitable for S. albula, yielding intermediate development time and higher survival relative to the other diets. Individualization of larvae favored S. albula development by producing overall greater weights of larvae and pupae, higher survival rates, and longer adult longevity. Cage size and number of couples per cage did not influence S. albula fecundity in the experiment conditions. Spodoptera albula can be satisfactorily reared on the artificial diet used for A. gemmatalis, using one larva per tube, and either density of adults at any cage size. Additional amendments are needed in the rearing methodology to achieve optimal conditions for larval development to adulthood.

Hematologic and serum biochemical data collected must be interpreted by comparison with normal reference intervals generated from healthy animals, within a similar population, because many blood parameters are influenced by diet, environment, and stress. Species-specific reference intervals for the endangered island fox (Urocyon littoralis) are not available. We reviewed hematology and serum biochemistry panels from 280 island foxes sampled from 1999-2008 and established normal reference intervals from clinically healthy foxes using a nonparametric approach. Blood parameters were analyzed for differences in age, sex, island of origin, and captivity status. Alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, and creatine kinase activities, as well as calcium and phosphorus concentrations, were significantly higher in juveniles than in adults, but total protein and globulin concentration was lower for juveniles than for adults. Lymphocyte and eosinophil counts, and blood urea nitrogen (BUN) concentration, in foxes from the northern Channel islands of California, USA (Santa Cruz, Santa Rosa, and San Miguel) were higher when compared with foxes from Santa Catalina Island to the south. Higher lymphocyte and eosinophil numbers in the northern island foxes may be associated with increased levels of parasitism on the northern islands. Differences in BUN concentration in both free-ranging and captive foxes may reflect dietary differences among islands. Although aggressive conservation programs have been enacted, island foxes are still susceptible to infectious and neoplastic diseases and, potentially, to toxins. Island fox species-specific reference intervals will enable managers and veterinarians to better care for sick and injured foxes and will contribute to future population health monitoring.

Griselinia littoralis, a native New Zealand shrub, was planted into a chronosequence (0 to 8 yrs since flooding) dominated by the non-indigenous shrub, Buddleja davidii in three New Zealand floodplains to determine to what extent facilitation and competitive inhibition may influe...

Drosophila littoralis is a latitudinally widespread European species of the Drosophila virilis group. The species has ample genetic variation in photoperiodism (adult diapause) and circadian rhythmicity (pupal eclosion rhythm), with adaptive latitudinal clines in both of them. The possible common genetic basis between the variability of photoperiodism and circadian rhythms was studied by a long-term crossing experiment. A northern strain (65 degrees N) having long critical day length (CDL = 19.9 h) for diapause, early phase of the entrained rhythm in LD 3:21 (psi(LD3:21) = 12.3 h), and short period (tau= 18.8 h) of the free-running rhythm for the eclosion rhythm was crossed with a southern strain (42 degrees N) having short CDL (12.4 h), late eclosion phase (psi(LD3:21) = 20.2 h), and long period (tau= 22.8 h). After 54 generations, including free recombination, artificial selection, and genetic drift, a novel strain resulted, having even more "southern" diapause and more "northern" eclosion rhythm characteristics than found in any of the geographical strains. The observed complete separation of eclosion rhythm characteristics from photoperiodism is a new finding in D. littoralis; in earlier studies followed for 16 generations, the changes had been mostly parallel. Evidently, the genes controlling the variability of the eclosion rhythm and photoperiodism in D. littoralis are different but closely linked. To test for the possible gene loci underlying the observed geographical variability, the period gene was studied in 10 strains covering all the known clock variability in D. littoralis. The authors sequenced the most suspected Thr-Gly region, which is known to take part in the adaptive clock variability in Drosophila melanogaster. No coding differences were found in the strains, showing that this region is not included in the adaptive clock variability in D. littoralis.

We characterized the dynamics of mortality factors affecting immature developmental stages of the fall armyworm, Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae). Multiple decrement life tables for egg and early larval stages of S. frugiperda in maize (Zea mays L.) fields were developed with and without augmentative releases of Telenomus remus Nixon (Hymenoptera: Platygastridae) from 2009 to 2011. Total egg mortality ranged from 73 to 81% and the greatest egg mortality was due to inviability, dislodgement, and predation. Parasitoids did not cause significant mortality in egg or early larval stages and the releases of T. remus did not increase egg mortality. Greater than 95% of early larvae died from predation, drowning, and dislodgment by rainfall. Total mortality due to these factors was largely irreplaceable. Results indicate that a greater effect in reducing generational survival may be achieved by adding mortality to the early larval stage of S. frugiperda. PMID:26098422

Many of the mechanisms underlying density-dependent regulation of populations, including contest competition and disease spread, depend on contact among neighboring animals. Understanding how variation in population density influences the frequency of contact among neighboring animals is therefore an important aspect to understanding the mechanisms underlying, and ecological consequences of, density-dependent regulation. However, contact rates are difficult to measure in the field and may be influenced by density through multiple pathways. This study explored how local density affects contact rates among Channel Island foxes (Urocyon littoralis) through two pathways: changes in home range size and changes in home range overlap. We tracked 40 radio-collared foxes at four sites on San Clemente Island, California. Fox densities at the four sites ranged from 2.8 ± 1.28 to 42.8 ± 9.43 foxes/km2. Higher fox densities were correlated with smaller home ranges (R2 = 0.526, F1,38 = 42.19, P

Foliar chemicals are variable within a plant and this may affect herbivore feeding preference. This study was carried out to quantify concentrations of primary (nitrogen, water, and total nonstructural carbohydrates) and secondary substances (sinigrin) in young and old leaves of Raphanus sativus L. and to evaluate performance and survival of a generalist herbivore Spodoptera litura F. feeding on them. Forty to 50-d-old R. sativus plants were used in both foliar chemical analysis and insect performance bioassays. Leaves located on the third to the sixth node from the base of the plant were defined as old leaves and the remaining leaves (from seventh node to the plant apex) of the plant were referred as young leaves in this study. All foliar chemicals except water differed significantly between young and old leaves. Moreover, young leaves were more nutritious but much more defended, based on sinigrin content, against S. litura than old leaves. Performance and survival of S. litura were reduced on young leaves as compared with old leaves. Male and female larval duration only differed significantly on young leaves. Female larval development time was longer than male development time on young leaves, but not on older leaves. Therefore, this study revealed that defenses in young leaves have differential effects upon male and female S. litura.

Freshwater mussels of the family Unionidae exhibit a particular form of mitochondria inheritance called double uniparental inheritance (DUI), in which the mitochondria are inherited by both male and female parents. The (M)ale and (F)emale mitogenomes are highly divergent within species. In the present study, we determine and describe the complete M and F mitogenomes of the Endangered freshwater mussel Potomida littoralis (Cuvier, 1798). The complete M and F mitogenomes sequences are 16 451 bp and 15 787 bp in length, respectively. Both F and M have the same gene content: 13 protein-coding genes (PCGs), 22 transfer RNA (trn) and 2 ribosomal RNA (rrn) genes. Bayesian analyses based on the concatenated nucleotide sequences of 12 PCGs and 2 rrn genes of both genomes, including mitogenome sequences available from related species, were performed. Male and Female lineages are monophyletic within the family, but reveal distinct phylogenetic relationships.

Freshwater mussels of the family Unionidae exhibit a particular form of mitochondria inheritance called double uniparental inheritance (DUI), in which the mitochondria are inherited by both male and female parents. The (M)ale and (F)emale mitogenomes are highly divergent within species. In the present study, we determine and describe the complete M and F mitogenomes of the Endangered freshwater mussel Potomida littoralis (Cuvier, 1798). The complete M and F mitogenomes sequences are 16 451 bp and 15 787 bp in length, respectively. Both F and M have the same gene content: 13 protein-coding genes (PCGs), 22 transfer RNA (trn) and 2 ribosomal RNA (rrn) genes. Bayesian analyses based on the concatenated nucleotide sequences of 12 PCGs and 2 rrn genes of both genomes, including mitogenome sequences available from related species, were performed. Male and Female lineages are monophyletic within the family, but reveal distinct phylogenetic relationships.

Freshwater mussels of the family Unionidae exhibit a particular form of mitochondria inheritance called double uniparental inheritance (DUI), in which the mitochondria are inherited by both male and female parents. The (M)ale and (F)emale mitogenomes are highly divergent within species. In the present study, we determine and describe the complete M and F mitogenomes of the Endangered freshwater mussel Potomida littoralis (Cuvier, 1798). The complete M and F mitogenomes sequences are 16 451 bp and 15 787 bp in length, respectively. Both F and M have the same gene content: 13 protein-coding genes (PCGs), 22 transfer RNA (trn) and 2 ribosomal RNA (rrn) genes. Bayesian analyses based on the concatenated nucleotide sequences of 12 PCGs and 2 rrn genes of both genomes, including mitogenome sequences available from related species, were performed. Male and Female lineages are monophyletic within the family, but reveal distinct phylogenetic relationships.

Successful xenotransfusion in an island fox ( Urocyon littoralis clementae) has not been previously reported but may be necessary in an emergency. An 11-yr-old male, intact, captive island fox was exhibiting clinical signs of rattlesnake envenomation including hypoperfusion, tachypnea, facial edema, and multifocal facial and cervical ecchymosis. Blood work revealed severe thrombocytopenia (18 K/μl) and anemia (Hct 15.8%). A presumptive diagnosis of rattlesnake ( Crotalus sp.) envenomation was made. Initial treatment included oxygen therapy, fluid therapy, antibiotics, antacids, pain medications, and polyvalent crotalid anti-venom. Emergency xenotransfusion using whole blood (45 ml) from a domestic dog was used due to worsening clinical signs from anemia. No acute or delayed transfusion reactions were observed in the fox and the patient made a full recovery 5 days later. Successful xenotransfusion in an island fox using whole blood from a domestic dog ( Canis lupus familiaris) is possible and may be lifesaving.

The utilization of storage lipids and their associated fatty acids (FA) is an important means for organisms to cope with periods of food shortage, however, little is known about the dynamics and FA mobilization in benthic copepods (order Harpacticoida). Furthermore, lipid depletion and FA mobilization may depend on the ambient temperature. Therefore, we subjected the temperate copepod Platychelipus littoralis to several intervals (3, 6 and 14 days) of food deprivation, under two temperatures in the range of the normal habitat temperature (4, 15°C) and under an elevated temperature (24°C), and studied the changes in FA composition of storage and membrane lipids. Although bulk depletion of storage FA occurred after a few days of food deprivation under 4°C and 15°C, copepod survival remained high during the experiment, suggesting the catabolization of other energy sources. Ambient temperature affected both the degree of FA depletion and the FA mobilization. In particular, storage FA were more exhausted and FA mobilization was more selective under 15°C compared with 4°C. In contrast, depletion of storage FA was limited under an elevated temperature, potentially due to a switch to partial anaerobiosis. Food deprivation induced selective DHA retention in the copepod’s membrane, under all temperatures. However, prolonged exposure to heat and nutritional stress eventually depleted DHA in the membranes, and potentially induced high copepod mortality. Storage lipids clearly played an important role in the short-term response of the copepod P. littoralis to food deprivation. However, under elevated temperature, the use of storage FA as an energy source is compromised. PMID:26986852

The utilization of storage lipids and their associated fatty acids (FA) is an important means for organisms to cope with periods of food shortage, however, little is known about the dynamics and FA mobilization in benthic copepods (order Harpacticoida). Furthermore, lipid depletion and FA mobilization may depend on the ambient temperature. Therefore, we subjected the temperate copepod Platychelipus littoralis to several intervals (3, 6 and 14 days) of food deprivation, under two temperatures in the range of the normal habitat temperature (4, 15 °C) and under an elevated temperature (24 °C), and studied the changes in FA composition of storage and membrane lipids. Although bulk depletion of storage FA occurred after a few days of food deprivation under 4 °C and 15 °C, copepod survival remained high during the experiment, suggesting the catabolization of other energy sources. Ambient temperature affected both the degree of FA depletion and the FA mobilization. In particular, storage FA were more exhausted and FA mobilization was more selective under 15 °C compared with 4 °C. In contrast, depletion of storage FA was limited under an elevated temperature, potentially due to a switch to partial anaerobiosis. Food deprivation induced selective DHA retention in the copepod's membrane, under all temperatures. However, prolonged exposure to heat and nutritional stress eventually depleted DHA in the membranes, and potentially induced high copepod mortality. Storage lipids clearly played an important role in the short-term response of the copepod P. littoralis to food deprivation. However, under elevated temperature, the use of storage FA as an energy source is compromised.

Nine of the most common lepidopteran active Cry proteins from Bacillus thuringiensis have been tested for activity against Spodoptera exigua. Because of possible intraspecific variability, three laboratory strains (FRA, HOL, and MUR) have been used. Mortality assays were performed with the three strains. LC(50) values for the active toxins were determined to the FRA and the HOL strains, whereas susceptibility of the MUR strain was assessed using only two concentrations. The results showed that Cry1Ca, Cry1Da, and Cry1Fa were the most effective toxins with all strains. Cry1Ab was found effective for the HOL strain, but very little effective against FRA (6.5-fold) and MUR strains. Cry1Aa and Cry1Ac were marginally toxic to all strains, whereas the rest of the toxins tested (Cry1Ba, Cry2Aa, and Cry2Ab) were non toxic. Significant differences in susceptibility among strains were also found for Cry1Da, being the FRA strain 25-fold more susceptible than the HOL strain. Growth inhibition, as an additional susceptibility parameter, was determined in the FRA strain with the 9 toxins. The toxicity profile obtained differed from that observed in mortality assays. Cry1Aa, Cry1Ab, Cry1Ac, Cry1Ca, Cry1Da, and Cry1Fa toxins produced a similar larval growth inhibition. Cry2Aa had a lower but clear effect on larval growth inhibition, whereas Cry1Ba and Cry2Ab did not have any effect.

Endophytic fungi, which live within host plant tissues without causing any visible symptom of disease, are important mediators of plant-herbivore interactions. These endophytes enhance resistance of host plant against insect herbivores mainly by productions of various alkaloid based defensive compounds in the plant tissue or through alterations of plant nutritional quality. Two endophytic fungi, i.e., Nigrospora sp. and Cladosporium sp., were isolated from Tinospora cordifolia (Thunb.) Miers, a traditional indian medicinal plant. Cauliflower (Brassica oleracea L.) plants were inoculated with these two endophytic fungi. The effect of endophyte infected and uninfected cauliflower plants were measured on the survival and development of Spodoptera litura (Fab.), a polyphagous pest. Endophyte infected cauliflower plants showed resistance to S. litura in the form of significant increase in larval and pupal mortality in both the fungi. Inhibitory effects of endophytic fungi also were observed on adult emergence, longevity, reproductive potential, as well as hatchability of eggs. Thus, it is concluded that antibiosis to S. litura could be imparted by artificial inoculation of endophytes and this could be used to develop alternative ecologically safe control strategies.

Melanism has long been thought to be a habitat adaptation with a fitness cost. Here we reported a homozygous melanic strain (SEM) of Spodoptera exigua (Hübner) (Insecta: Lepidoptera: Noctuidae) established with black pupae spontaneously occurring within a typical laboratory population (SEW). The melanization is expressed globally, and only in the pupal stage. After pupation, the melanic SEM pupae gradually accumulate melanin to become completely black within 6 hours, whereas the wild-type SEW pupae gradually turn yellow-brown. The melanic SEM strain exhibits faster development in all life stages, heavier pupa weight, more mating time, higher fecundity, and accordingly, higher net reproductive rate and population trend index. While no reproductive isolation was observed between the SEM and SEW strains, the mating times per female of the reciprocal crosses and the SEM intracrosses were significantly higher than those of the SEW intracrosses. This represents a rare case of melanization that has fitness gains, rather than costs. Analysis of the life-history traits of this case and 14 previously reported cases of insect melanism indicate that none of melanization origin, stage, space and variation type determining whether melanism will cause fitness gain or cost. PMID:26039886

Digestive and detoxification enzyme activity and nutrient composition were examined in the body of fourth instar beet armyworms, Spodoptera exigua (Hübner), fed on transgenic Bacillus thuringiensis (Bt) and non-Bt cotton for different time periods. Nutrient composition and specific enzyme activities differed significantly between the S. exigua fed Bt vs. non-Bt cotton. At 1, 6 and 24 h, free fatty acid and glucose levels were significantly lower in S. exigua fed on Bt cotton than those fed on non-Bt cotton. S. exigua fed on Bt cotton had significantly higher trypsin and total superoxide dismutase (T-SOD) activities and significantly lower lipase, carboxylesterase and acetylcholinesterase activities than non-Bt fed worms for all feeding time periods. Differences were also observed among feeding times within each cotton variety group. Significantly lower free fatty acid and total amino acid were observed in S. exigua fed on Bt cotton for 24 h than in those fed for 1 h. Significantly lower activities of lipase and trypsin were detected in S. exigua fed on Bt cotton for 24 h than those for 1 and 4 h. However, carboxylesterase and acetylcholinesterase activities in S. exigua fed on Bt cotton for 24 h were significantly higher than those for 1, 4 and 6 h. The interaction between cotton variety and feeding time significantly affected the activities of lipase, trypsin, acetylcholinesterase and T-SOD enzymes in S. exigua. Measuring the temporal allocation of protection and detoxification enzyme activities in the body of S. exigua in response to B. thuringiensis can provide a meaningful evaluation on the metabolic tolerance of herbivorous insects under the continuous selection pressure of a toxic protein.

The presence of lignin within biomass impedes the production of liquid fuels. Plants with altered lignin content and composition are more amenable to lignocellulosic conversion to ethanol and other biofuels but may be more susceptible to insect damage where lignin is an important resistance factor. However, reduced lignin lines of switchgrasses still retained insect resistance in prior studies. Therefore, we hypothesized that sorghum lines with lowered lignin content will also retain insect resistance. Sorghum excised leaves and stalk pith Sorghum bicolor (L.) Moench (Poales: Poaceae) from near isogenic brown midrib (bmr) 6 and 12 mutants lines, which have lowered lignin content and increased lignocellulosic ethanol conversion efficiency, were examined for insect resistance relative to wild-type (normal BTx623). Greenhouse and growth chamber grown plant tissues were fed to first-instar larvae of corn earworms, Helicoverpa zea (Boddie) and fall armyworms Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae), two sorghum major pests. Younger bmr leaves had significantly greater feeding damage in some assays than wild-type leaves, but older bmr6 leaves generally had significantly less damage than wild-type leaves. Caterpillars feeding on the bmr6 leaves often weighed significantly less than those feeding on wild-type leaves, especially in the S. frugiperda assays. Larvae fed the pith from bmr stalks had significantly higher mortality compared with those larvae fed on wild-type pith, which suggested that bmr pith was more toxic. Thus, reducing lignin content or changing subunit composition of bioenergy grasses does not necessarily increase their susceptibility to insects and may result in increased resistance, which would contribute to sustainable production. PMID:25601946

Significant changes usually take place in the internal metabolism of insects during metamorphosis. The glycolysis-tricarboxylic acid (glycolysis-TCA) pathway is important for energy metabolism. To elucidate its dynamics, the mRNA levels of genes involved in this pathway were examined in the midgut of Spodoptera litura during metamorphosis, and the pyruvate content was quantified. The expression patterns of these genes in response to starvation were examined, and the interaction between protein phosphatase 1 (PP1) and phosphofructokinase (PFK) was studied. The results revealed that the expression or activities of most glycolytic enzymes was down-regulated in prepupae and then recovered in some degree in pupae, and all TCA-related genes were remarkably suppressed in both the prepupae and pupae. Pyruvate was enriched in the pupal midgut. Taken together, these results suggest that insects decrease both glycolysis and TCA in prepupae to save energy and then up-regulate glycolysis but down-regulate TCA in pupae to increase the supply of intermediates for construction of new organs. The expression of all these genes were down-regulated by starvation, indicating that non-feeding during metamorphosis may be a regulator of glycolysis-TCA pathway in the midgut. Importantly, interaction between PP1 and PFK was identified and is suggested to be involved in the regulation of glycolysis.

Beet armyworm, Spodoptera exigua (Huebner), larvae were placed on sugarbeet (Beta vulgaris L.) and pigweed (Amaranthus hybridus L.) plants in outdoor chambers in which the plants were growing at either the ambient ([approximately] 350 [mu]l liter[sup [minus]1]) or ambient plus 350 [mu]l liter[sup [minus]1] ([approximately] 700 [mu]l liter[sup [minus]1]) carbon dioxide concentration. A series of experiments was performed to determine if larvae reduced plant growth differently at the two carbon dioxide concentrations in either species and if the insect growth or survival differed with carbon dioxide concentration. Leaf nitrogen, water, starch, and soluble carbohydrate contents were measured to assess carbon dioxide concentration effects on leaf quality. Insect feeding significantly reduced plant growth in sugarbeet plants at 350 [mu]l liter[sup [minus]1] but not at 700 [mu]l liter[sup [minus]1] nor in pigweed at either carbon dioxide concentration. Larval survival was greater on sugarbeet plants at the elevated carbon dioxide concentration. Increased survival occurred only if the insects were at the elevated carbon dioxide concentration and consumed leaf material grown at the elevated concentration. Leaf quality was only marginally affected by growth at elevated carbon dioxide concentration in these experiments. The results indicate that in designing experiments to predict effects of elevated atmospheric carbon dioxide concentrations on plant-insect interactions, both plants and insects should be exposed to the experimental carbon dioxide concentrations, as well as to as realistic environmental conditions as possible.

Beet armyworm, Spodoptera exigua, is a major pest of cotton around the world. With the increase of resistance to Bacillus thuringiensis (Bt) toxin in transgenic cotton plants, there is a need to develop an alternative control approach that can be used in combination with Bt transgenic crops as part of resistance management strategies. MicroRNAs (miRNAs), a non-coding small RNA family (18-25 nt), play crucial roles in various biological processes and over-expression of miRNAs has been shown to interfere with the normal development of insects. In this study, we identified 127 conserved miRNAs in S. exigua by using small RNA deep sequencing technology. From this, we tested the effects of 11 miRNAs on larval development. We found three miRNAs, Sex-miR-10-1a, Sex-miR-4924, and Sex-miR-9, to be differentially expressed during larval stages of S. exigua. Oral feeding experiments using synthetic miRNA mimics of Sex-miR-10-1a, Sex-miR-4924, and Sex-miR-9 resulted in suppressed growth of S. exigua and mortality. Over-expression of Sex-miR-4924 caused a significant reduction in the expression level of chitinase 1 and caused abortive molting in the insects. Therefore, we demonstrated a novel approach of using miRNA mimics to control S. exigua development.

Background Daucus littoralis Smith subsp. hyrcanicus Rech.f. (Apiaceae) is an endemic species in northern parts of Iran where it is commonly named Caspian carrot. The fruits have been used as condiment. Methods In a series of in vitro assays, antioxidant (DPPH and FRAP assays), cytotoxic and antimicrobial activities of different extracts of roots and fruits were evaluated for the first time. The separation and purification of the compounds were carried out on the most potent extracts using various chromatographic methods and identified by spectroscopic data (1H and 13C NMR). Results The results showed that among the extracts only fruit methanol extract (FME) has significant antioxidant activity (IC50 = 145.93 μg.ml-1 in DPPH assay and 358 ± 0.02 mmol FeII/g dry extract in FRAP assay). The radical scavenging activity of FME at 400 μg.ml-1 was comparable with α-tocopherol (40 μg.ml-1) and with BHA (100 μg.ml-1) (p > 0.05). FME did not show any toxicity against cancerous and normal cell lines. Fruit ethyl acetate extract (FEE) had cytotoxic activity against breast carcinoma and hepatocellular carcinoma cells (IC50 168.4 and 185 μg.ml-1, respectively), while it did not possess antioxidant activity in comparison with α-tocopherol and BHA as standard compounds. Ethyl acetate and methanol extract of fruits showed antimicrobial activity against Staphylococcus aureus (MIC: 3.75 mg.ml-1) and Candida albicans (MIC: 15.6 and 7.8 mg.ml-1, respectively). Four terpenoids were isolated form FEE including: β-sitosterol (1), stigmasterol (2), caryophyllene oxide (3), β-amyrin (4). Also, three flavonoids namely quercetin 3-O-β-glucoside (5), quercetin 3-O-β-galactoside (6) and luteolin (7) were isolated from FME. Conclusion This study showed that FEE and FME of D. littoralis Smith subsp. hyrcanicus Rech.f. had the highest biological activities which may be correlated with in vitro cytotoxic, antimicrobial and antioxidant activities of terpenoids and

In this work, we investigate the physiological responses to P deficiency (5μM KH2PO4=D), salt stress (400mM NaCl=C+S), and their combination (D+S) on the facultative halophyte Aeluropus littoralis to understand how plants adapt to these combined stresses. When individually applied, both P deficiency and salinity significantly restricted whole plant growth, with a more marked effect of the latter stress. However, the effects of the two stresses were not additive in plant biomass production since the response of plants to combined salinity and P deficiency was similar to that of plants grown under salt stress alone. In addition the observed features under salinity alone are kept when plants are simultaneously subjected to the combined effects of salinity and P deficiency such as biomass partitioning; the synthesis of proline and the K(+)/Na(+) selectivity ratio. Thus, increasing P availability under saline conditions has no significant effect on salt tolerance in this species. Plants cultivated under the combined effects of salinity and P deficiency exhibited the lowest leaf water potential. This trend was associated with a high accumulation of Na(+), Cl(-) and proline in shoots of salt treated plants suggesting the involvement of these solutes in osmotic adjustment. Proline could be involved in other physiological processes such as free radical scavenging. Furthermore, salinity has no significant effect on phosphorus acquisition when combined with a low P supply and it significantly decreased this parameter when combined with a sufficient P supply. This fact was probably due to salt's effect on P transporters. In addition, shoot soluble sugars accumulation under both P deficiency treatments with and without salt likely play an important role in the adaptation of A. littoralis plants to P shortage applied alone or combined with salinity. Moreover, there was a strong correlation between shoot and root intracellular acid phosphatase activity and phosphorus use

A PCR-based method was used to identify and distinguish among 40 uncharacterized nucleopolyhedrovirus (NPV) isolates from the moth Spodoptera frugiperda that were part of an insect virus collection. Phylogenetic analysis was carried out with sequences amplified from two strongly conserved loci (pol...

Since the first fatty acid amino acid conjugate (FAC) was isolated from regurgitant of Spodoptera exigua larvae in 1997 [volicitin: N-(17-hydroxylinolenoyl)- L-glutamine], their role as elicitors of induced responses in plants has been well documented. However, studies of the biosyntheses as well as...

The pheromone lure for the exotic species Spodoptera exempta was successful at attracting two native species, S. latifascia and S. albula. Trapping was conducted in north-central Florida and in southern Texas. Large numbers of both native species were collected throughout the season....

Three biopesticide parameters were evaluated for a fast-killing isolate (3AP2) Spodoptera frugiperda multiple nucleopolyhedrovirus (SfMNPV) and a wild-type isolate (Sf3) of the same baculovirus. Both isolates were evaluated for virus production using in vivo methods, for speed of kill based on bioas...

Feeding by insect herbivores activates plant signaling pathways, resulting in the enhanced production of secondary metabolites and other resistance-related traits by injured plants. These traits can reduce insect fitness, deter feeding, and attract beneficial insects. Organic and inorganic chemicals applied as a foliar spray, seed treatment, or soil drench can activate these plant responses. Azelaic acid (AA), benzothiadiazole (BTH), gibberellic acid (GA), harpin, and jasmonic acid (JA) are thought to directly mediate plant responses to pathogens and herbivores or to mimic compounds that do. The effects of these potential elicitors on the induction of plant defenses were determined by measuring the weight gains of fall armyworm, Spodoptera frugiperda (J. E. Smith) (FAW) (Lepidoptera: Noctuidae) larvae on four crop plants, cotton, corn, rice, and soybean, treated with the compounds under greenhouse conditions. Treatment with JA consistently reduced growth of FAW reared on treated cotton and soybean. In contrast, FAW fed BTH- and harpin-treated cotton and soybean tissue gained more weight than those fed control leaf tissue, consistent with negative crosstalk between the salicylic acid and JA signaling pathways. No induction or inconsistent induction of resistance was observed in corn and rice. Follow-up experiments showed that the co-application of adjuvants with JA failed to increase the effectiveness of induction by JA and that soybean looper [Chrysodeixis includens (Walker)], a relative specialist on legumes, was less affected by JA-induced responses in soybean than was the polyphagous FAW. Overall, the results of these experiments demonstrate that the effectiveness of elicitors as a management tactic will depend strongly on the identities of the crop, the pest, and the elicitor involved. PMID:26332833

Ear mites (Otodectes cynotis) and ear canal tumors are highly prevalent among federally endangered Island foxes (Urocyon littoralis catalinae) living on Santa Catalina Island off the coast of Southern California. Since studies began in the 1990s, nearly all foxes examined were found to be infected with ear mites, and ceruminous gland tumors (carcinomas and adenomas) were detected in approximately half of all foxes ≥ 4 years of age. We hypothesized that reduction of ear mite infection would reduce otitis externa and ceruminous gland hyperplasia, a risk factor for tumor development. In this study, we conducted a randomized field trial to assess the impact of acaricide treatment on ear mite prevalence and intensity of infection, otitis externa, ceruminous gland hyperplasia, and mite-specific IgG and IgE antibody levels. Treatment was highly effective at eliminating mites and reducing otitis externa and ceruminous gland hyperplasia, and mite-specific IgG antibody levels were significantly lower among uninfected foxes. Ceruminous gland hyperplasia increased in the chronically infected, untreated foxes during the six month study. Our results provide compelling evidence that acaricide treatment is an effective means of reducing ear mites, and that mite removal in turn reduces ear lesions and mite-specific IgG antibody levels in Santa Catalina Island foxes. This study has advanced our understanding of the underlying pathogenesis which results in ceruminous gland tumors, and has helped inform management decisions that impact species conservation. PMID:26641820

Prey response to novel predators influences the impacts on prey populations of introduced predators, bio-control efforts, and predator range expansion. Predicting the impacts of novel predators on native prey requires an understanding of both predator avoidance strategies and their potential to reduce predation risk. We examine the response of island foxes (Urocyon littoralis) to invasion by golden eagles (Aquila chrysaetos). Foxes reduced daytime activity and increased night time activity relative to eagle-naïve foxes. Individual foxes reverted toward diurnal tendencies following eagle removal efforts. We quantified the potential population impact of reduced diurnality by modeling island fox population dynamics. Our model predicted an annual population decline similar to what was observed following golden eagle invasion and predicted that the observed 11% reduction in daytime activity would not reduce predation risk sufficiently to reduce extinction risk. The limited effect of this behaviorally plastic predator avoidance strategy highlights the importance of linking behavioral change to population dynamics for predicting the impact of novel predators on resident prey populations.

The island fox (Urocyon littoralis catalinae) population on Santa Catalina Island, California, USA declined precipitously in 1999 with an approximate 95% reduction on their eastern range, an area representing 87% of the island. During this investigation, between October 1999 and April 2000, evidence of live foxes dramatically decreased. The only carcass recovered during the decline succumbed to a co-infection of canine distemper virus (CDV) and toxoplasmosis. Sequence analysis of the viral P gene, derived by polymerase chain reaction, indicated that the virus was closely related to CDV from a mainland USA raccoon (Procyon lotor). Nine of 10 foxes trapped in 1999-2000, on the eastern portion of the island after the decline, had serologic evidence of exposure to CDV, whereas only four of 19 foxes trapped in this region in 1998 had antibodies reactive against CDV. The confirmation of CDV in one deceased fox, evidence of exposure to CDV in east-end foxes in 1999-2000 compared to 1998, and documentation of raccoon introductions to the island, implicates canine distemper as the cause of the population decline.

Ear mites (Otodectes cynotis) and ear canal tumors are highly prevalent among federally endangered Island foxes (Urocyon littoralis catalinae) living on Santa Catalina Island off the coast of Southern California. Since studies began in the 1990s, nearly all foxes examined were found to be infected with ear mites, and ceruminous gland tumors (carcinomas and adenomas) were detected in approximately half of all foxes ≥ 4 years of age. We hypothesized that reduction of ear mite infection would reduce otitis externa and ceruminous gland hyperplasia, a risk factor for tumor development. In this study, we conducted a randomized field trial to assess the impact of acaricide treatment on ear mite prevalence and intensity of infection, otitis externa, ceruminous gland hyperplasia, and mite-specific IgG and IgE antibody levels. Treatment was highly effective at eliminating mites and reducing otitis externa and ceruminous gland hyperplasia, and mite-specific IgG antibody levels were significantly lower among uninfected foxes. Ceruminous gland hyperplasia increased in the chronically infected, untreated foxes during the six month study. Our results provide compelling evidence that acaricide treatment is an effective means of reducing ear mites, and that mite removal in turn reduces ear lesions and mite-specific IgG antibody levels in Santa Catalina Island foxes. This study has advanced our understanding of the underlying pathogenesis which results in ceruminous gland tumors, and has helped inform management decisions that impact species conservation.

Eicosanoids mediate cellular and humoral immune responses in the beet armyworm, Spodoptera exigua, including activation of prophenoloxidase (PPO). PPO activation begins with release of its inactive zymogen, PPO, from oenocytoids in response to prostaglandins (PGs). Based on the biomedical literatur...

The benefits, equipment used, commercially available sources, and the indications and techniques for administration of enteral nutrients are reviewed. In many malabsorption states, enteral feeding is preferable and parenteral nutrients are seldom indicated. Transitional enteral nutrient support usually is indicated after parenteral nutrient therapy. Enteral tube-feeding formulas should be matched to the patient's needs; formulas using blenderized natural foods or intact isolated nutrients are appropriate for patients with intact gastrointestinal tracts. Patients should be monitored for glucosuria and hyperglycemia, bloating, nausea, dehydration, and renal, hepatic and hematologic status. Formula dilution, and a reduced flow rate or use of continuous-drip feeding, will reduce the incidence of osmotic diarrhea. The effectiveness, low cost and low potential for serious complications make enteral feeding preferable to parenteral nutrient therapy for many patients.

Background Recovering endangered species would benefit from identifying and ranking of the factors that threaten them. Simply managing for multiple positive influences will often aid in recovery; however, the relative impacts of multiple threats and/or interactions among them are not always predictable. We used a series of experiments and quantitative observational studies to examine the importance of five potential limiting factors to the abundance of a state-listed endangered hemiparasitic annual forb, Cordylanthus rigidus ssp. littoralis (C.r.l., California, USA): host availability, mammalian herbivores, insect seed predators, fire suppression, and exotic species. While this initial assessment is certainly not a complete list, these factors stem from direct observation and can inform provisional recommendations for management and further research. Methodology and Principal Findings Studies were conducted at five sites and included assessments of the influence of host availability, exotic species, exclusion of mammalian herbivores and insect seed predators on C.r.l. productivity, and simulated effects of fire on seed germination. C.r.l. was limited by multiple threats: individuals with access to host species produced up to three times more inflorescences than those lacking hosts, mammalian herbivory reduced C.r.l. size and fecundity by more than 50% and moth larvae reduced seed production by up to 40%. Litter deposition and competition from exotic plant species also appears to work in conjunction with other factors to limit C.r.l. throughout its life cycle. Conclusions and Significance The work reported here highlights the contribution that a series of small-scale studies can make to conservation and restoration. Taken as a whole, the results can be used immediately to inform current management and species recovery strategies. Recovery of C.r.l. will require management that addresses competition with exotic plant species, herbivore pressure, and availability of

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The aim of this study was to evaluate the effects of silicon application and administration of the phytohormone gibberellic acid on resistance of the corn plants to the fall armyworm (FAW), Spodoptera frugiperda, and their vegetative characteristics. We evaluated larval and pupal duration, survival and biomass, and adult longevity, malformation and fecundity of S. frugiperda after feeding on plant matter treated with silicon and/or gibberellic acid. The feeding preference of FAW first-instar larvae, the total leaf area consumed by the insects, and the vegetative parameters of corn plants were also evaluated. No significant differences were observed in the measured parameters of larval and pupal stages of S. frugiperda in response to silicon or gibberellic acid. In adult stage insects, the number of eggs per female was significantly reduced in insects derived from larvae fed plants treated with silicon or gibberellic acid. In a non-preference test, 48 h after release, caterpillars preferred control untreated plants and consumed less matter from plants that had received hormonal treatment (gibberellic acid). Gibberellic acid also altered the vegetative characteristics of plants, by increasing their height, shoot fresh and dry mass, and silicon content. We conclude that gibberellic acid can alter the vegetative characteristics and silicon uptake of corn plants, leading to a reduction in their consumption by S. frugiperda larvae and a decrease in female insect oviposition.

Vip3 proteins have been described to be secreted by Bacillus thuringiensis during the vegetative growth phase and to display a broad insecticidal spectrum against lepidopteran larvae. Vip3Aa protoxin has been reported to be significantly more toxic to Spodoptera frugiperda than to Spodoptera exigua and differences in the midgut processing have been proposed to be responsible. In contrast, we have found that Vip3Ae is essentially equally toxic against these two species. Proteolysis experiments were performed to study the stability of Vip3A proteins to peptidase digestion and to see whether the differences found could explain differences in toxicity against these two Spodoptera species. It was found that activation of the protoxin form and degradation of the 62kDa band took place at lower concentrations of trypsin when using Vip3Aa than when using Vip3Ae. The opposite effect was observed for chymotrypsin. Vip3Aa and Vip3Ae protoxins were effectively processed by midgut content extracts from the two Spodoptera species and the proteolytic activation did not produce a peptidase resistant core under these in vitro conditions. Digestion experiments performed with S. frugiperda chromatography-purified digestive serine peptidases showed that the degradation of the Vip3A toxins active core is mainly due to the action of cationic chymotrypsin-like peptidase. Although the digestion patterns of Vip3A proteins do not always correlate with toxicity, the peptidase stability of the 62kDa core is in agreement with intraspecific differences of toxicity of the Vip3Aa protein.

Nickel is an environmental pollutant that adversely affects the male reproductive system. In the present study, the effects of nickel exposure on Spodoptera litura Fabricius were investigated by feeding larvae artificial diets containing different doses of nickel for three generations. Damage to testes and effects on male reproduction were examined. The amount of nickel that accumulated in the testes of newly emerged males increased as the nickel dose in the diet increased during a single generation. Nickel exposure increased the amount of thiobarbituric acid reactive substances and decreased the amount of glutathione in treatment groups compared with the control. The activity levels of the antioxidant response indices superoxide dismutases, catalase, and glutathione peroxidase in the testes showed variable dose-dependent relationships with nickel doses and duration of exposure. Nickel doses also disrupted the development of the testes by decreasing the weight and volume of testes and the number of eupyrene and apyrene sperm bundles in treatment groups compared with the control. When the nickel-treated males mated with normal females, fecundity was inhibited by the higher nickel doses in all three generations, but fecundity significantly increased during the second generation, which received 5 mg kg(-1) nickel. Hatching rates in all treatments significantly decreased in a dose-dependent manner in the three successive generations. The effects of nickel on these parameters correlated with the duration of nickel exposure. Results indicate assays of testes may be a novel and efficient means of evaluating the effects of heavy metals on phytophagous insects in an agricultural environment.

The brown alga Pilayella littoralis was used as a new biosorbent in an on-line metal preconcentration procedure in a flow-injection system. Al, Co, Cu and Fe were determined in lake water samples by inductively coupled plasma optical emission spectrometry (ICP-OES) after preconcentration in a silica-immobilized alga column. Like other algae, P. littoralis exhibited strong affinity for these metals proving to be an effective accumulation medium. Metals were bound at pH 5.5 and were displaced at pH<2 with diluted HCl. The enrichment factors for Cu(II), Fe(III), Al(III) and Co(II) were 13, 7, 16 and 11, respectively. Metal sorption efficiency ranged from 86 to 90%. The method accuracy was assessed by using drinking water certified reference material and graphite furnace atomic absorption spectrometry (GFAAS) as a comparison technique. The column procedure allowed a less time consuming, easy regeneration of the biomaterial and rigidity of the alga provided by its immobilization on silica gel.

This set of documents consists of English, French, and Spanish translations of four pamphlets on breast-feeding. The pamphlets provide information designed for lay persons, academics and professionals, health personnel and educators, and policy-makers. The contents cover health-related differences between breast and bottle milk; patterns of…

This module on tube feedings is intended for use in inservice or continuing education programs for persons who work in long-term care. Instructor information, including teaching suggestions and a listing of recommended audiovisual materials and their sources appear first. The module goal and objectives are then provided. A brief discussion follows…

Adult nematode parasites were recovered from the heart blood of a deceased island fox (Urocyon littoralis) submitted for necropsy to determine the cause of death. Examination of the recovered nematodes supported the generic diagnosis of Angiocaulus, a parasite in Angiostrongylidae found in domestic and wild canids and mustelids. Specific diagnosis of the worms from the island fox as Angiocaulus gubernaculatus is based on the morphology of the dorsal ray in the copulatory bursa of the male worm and its comparison with published descriptions of Angiocaulus raillieti and A. gubernaculatus. Although A. gubernaculatus has been typically associated with mustelid hosts, its occurrence in the island fox indicates that the host distribution for the parasite may not be as restricted as previously believed.

We identified a reactive natural toxin, biliatresone, from Dysphania glomulifera and D. littoralis collected in Australia that produces extrahepatic biliary atresia in a zebrafish model. Three additional isoflavonoids, including the known isoflavone betavulgarin, were also isolated. Biliatresone is in the very rare 1,2-diaryl-2-propenone class of isoflavonoids. The α-methylene of the 1,2-diaryl-2-propenone of biliatresone spontaneously reacts via Michael addition in the formation of water and methanol adducts. The lethal dose of biliatresone in a zebrafish assay was 1 μg/mL, while the lethal dose of synthetic 1,2-diaryl-2-propen-1-one was 5 μg/mL, suggesting 1,2-diaryl-2-propenone as the toxic Michael acceptor. PMID:26175131

We identified a reactive natural toxin, biliatresone, from Dysphania glomulifera and D. littoralis collected in Australia that produces extrahepatic biliary atresia in a zebrafish model. Three additional isoflavonoids, including the known isoflavone betavulgarin, were also isolated. Biliatresone is in the very rare 1,2-diaryl-2-propenone class of isoflavonoids. The α-methylene of the 1,2-diaryl-2-propenone of biliatresone spontaneously reacts via Michael addition in the formation of water and methanol adducts. The lethal dose of biliatresone in a zebrafish assay was 1 μg/mL, while the lethal dose of synthetic 1,2-diaryl-2-propen-1-one was 5 μg/mL, suggesting 1,2-diaryl-2-propenone as the toxic Michael acceptor.

Understanding how human activities have influenced the foraging ecology of wildlife is important as our planet faces ongoing and impending habitat and climatic change. We review the canine surrogacy approach (CSA)-a tool for comparing human, dog, and other canid diets in the past-and apply CSA to investigate possible ancient human resource provisioning in an endangered canid, the California Channel Islands fox (Urocyon littoralis). We conducted stable isotope analysis of bone collagen samples from ancient and modern island foxes (n = 214) and mainland gray foxes (Urocyon cinereoargenteus, n = 24). We compare these data to isotope values of ancient humans and dogs, and synthesize 29 Accelerator Mass Spectrometry (AMS) radiocarbon dates that fine-tune the chronology of island foxes. AMS dates confirm that island foxes likely arrived during the early Holocene (>7300 cal BP) on the northern islands in the archipelago and during the middle Holocene (>5500 cal BP) on the southern islands. We found no evidence that island foxes were consistently using anthropogenic resources (e.g., food obtained by scavenging around human habitation sites or direct provisioning by Native Americans), except for a few individuals on San Nicolas Island and possibly on San Clemente and Santa Rosa islands. Decreases in U. littoralis carbon and nitrogen isotope values between prehistoric times and the 19th century on San Nicolas Island suggest that changes in human land use from Native American hunter-gatherer occupations to historical ranching had a strong influence on fox diet. Island foxes exhibit considerable dietary variation through time and between islands and have adapted to a wide variety of climatic and cultural changes over the last 7300 years. This generalist foraging strategy suggests that endemic island foxes may be resilient to future changes in resource availability.

Background. Calomera littoralis is a Palearctic species, widely distributed in Europe; inhabiting predominantly its Atlantic, Mediterranean and Black Sea coastlines. Methods. Its phylogeography on the Balkan Peninsula and on the north-western Black Sea coast was inferred using a 697 bp long portion of the mitochondrial COI gene, amplified from 169 individuals collected on 43 localities. Results. The results revealed two genetically divergent groups/lineages, the southern one inhabiting both the Balkan Peninsula and the Pontic Region and the northern one found exclusively in the Pontic Region. Species delimitation based on DNA barcoding gap suggested an interspecific level of divergence between these groups. Multivariate analysis of eight male and female morphometric traits detected no difference between the groups, implying they may represent cryptic species. The Bayesian time-calibrated reconstruction of phylogeny suggested that the lineages diverged ca. 2.3 Ma, in early Pleistocene. Discussion. The presence of the two genetically divergent groups results most likely from contemporary isolation of the Pontic basin from the Mediterranean that broke the continuous strip of coastal habitats inhabited by C. littoralis. Demographic analyses indicated that both lineages have been in demographic and spatial expansion since ca. 0.15 Ma. It coincides with the terminal stage of MIS-6, i.e., Wartanian/Saalian glaciation, and beginning of MIS-5e, i.e., Eemian interglacial, during which, due to eustatic sea level rise, a wide connection between Mediterranean and the Pontic basin was re-established. This, along with re-appearance of coastal habitats could initiate north-east expansion of the southern lineage and its secondary contact with the northern one. The isolation of the Pontic basin from the Mediterranean during the Weichselian glaciation most likely did not have any effect on their phylogeography. PMID:27547517

Biotransformation of alpha-terpineol by the common cutworm (Spodoptera litura) larvae was investigated. alpha-Terpineol was mixed in an artificial diet, and the diet was fed to the larvae (fourth-fifth instar) of S. litura. Metabolites were isolated from the frass and analyzed spectroscopically. Main metabolites were 7-hydroxy-alpha-terpineol (p-menth-1-ene-7,8-diol) and oleuropeic acid (8-hydroxy-p-menth-1-en-7-oic acid). Intestinal bacteria from the frass of larvae did not participate in the metabolism of alpha-terpineol. alpha-Terpineol was preferentially oxidized at the C-7 position (allylic methyl group) by S. litura larvae.

Plants synthesize specialized metabolites which possess extremely important ecological functions including direct defense, indirect defense, and signaling. The optimal defense theory (ODT) proposes that defensive metabolites are preferentially allocated to the tissues with high fitness value or in locations that are easily injured. In our present study, using the model plant Nicotiana benthamiana, we found that direct defense of N. benthamiana against Spodoptera litura (Fabricius) larvae showed spatial differences in the sites producing defensive chemicals. The upper leaves possessed significantly stronger direct defense ability than the middle and lower leaves. Interestingly, the strong defense ability of the upper leaves was not due to occurrences of well-known defensive metabolites such as nicotine and chlorogenic acid. After damage, the middle and lower leaves emitted higher amounts of (Z)-3-hexen-1-ol than the upper leaves, which could both attract larvae and significantly increase the amount of middle and lower leaf eaten by the larvae. The spatial difference in (Z)-3-hexen-1-ol emission may be due to spatial differences in expression of lipoxygenase (NbLOX2), which is responsible for the formation and emission of (Z)-3-hexen-1-ol. This study provided new insight into ODT, showing that plants effectively protect easily injured tissues through reduction in concentration of herbivore-feeding stimulant in the tissues.

The stomatogastric nervous system plays a pivotal role in feeding behaviour. Central to this system is the frontal ganglion, which is responsible for foregut motor activity, and hence the passage of food through the gut. Many insect peptides, which exhibit myoactivity on the visceral muscles of the gut in vitro, have been detected in the stomatogastric nervous system by immunochemical or mass spectrometric techniques. This localisation of myoactive peptides, particularly in the frontal ganglion, implies roles for these peptides in the neural control and modulation of feeding in insects. Insect sulfakinins, tachykinins, allatotropin and proctolin have all been shown to stimulate the foregut muscles, whereas myosuppressins, myoinhibitory peptides and allatostatins all inhibited spontaneous contractions of the foregut in a variety of insects. Some of these peptides, when injected, inhibited feeding in vivo. Both the A-type and B-type allatostatins suppressed feeding activity when injected into the cockroach, Blattella germanica and the Manduca sexta C-type allatostatin and allatotropin inhibited feeding when injected into the larvae of two noctuid moths, Lacanobia oleracea and Spodoptera frugiperda, respectively. Injection of sulfakinins into the fly Phormia regina, the locust Schistocera gregaria and the cockroach B. germanica also suppressed feeding, whereas silencing the sulfakinin gene through the injection of double stranded RNA resulted in an increase in food consumption in the cricket Gryllus bimaculatus. The regulation of feeding in insects is clearly very complex, and involves the interaction of a number of mechanisms, one of which is the release, either centrally or locally, of neuropeptides. However, the role of neuropeptides, their mechanisms of action, interactions with each other, and their release are still poorly understood. It is also unclear why insects possess such a number of different peptides, some with multiples copies or homologues, which

Four different cactus species growing in the United States, Stenocereus thurberi growing in Organ Pipe Cactus National Monument, Arizona, Opuntia littoralis and Opuntia ficus-indica, growing on Santa Catalina Island, California, and Opuntia stricta, growing in northern Alabama, were examined for the presence of silica bodies (opaline phytoliths). Silica bodies were found in all four of these cactus species, parallelepiped-shaped crystals in S. thurberi, and starburst-shaped crystalline structures in the three Opuntia species. In addition, the essential oils of the four cactus species were obtained by hydrodistillation and analyzed by gas chromatography - mass spectrometry. To our surprise, S. thurberi, O. littoralis, and O. ficus-indica (but not O. stricta) essential oils contained cyclic oligosiloxanes. To our knowledge, cyclic oligosiloxanes have not been previously found as essential oil components.

The corn- and rice-strains of Spodoptera frugiperda exhibit several genetic and behavioral differences and appear to be undergoing ecological speciation in sympatry. Previous studies reported conflicting results when investigating male attraction to pheromone lures in different regions, but this cou...

... virus; exemption from the requirement of a tolerance. 180.1118 Section 180.1118 Protection of... polyhedrosis virus; exemption from the requirement of a tolerance. An exemption from the requirement of a tolerance is established for the microbial pest control agent Spodoptera exigua nuclear polyhedrosis...

... virus; exemption from the requirement of a tolerance. 180.1118 Section 180.1118 Protection of... polyhedrosis virus; exemption from the requirement of a tolerance. An exemption from the requirement of a tolerance is established for the microbial pest control agent Spodoptera exigua nuclear polyhedrosis...

... virus; exemption from the requirement of a tolerance. 180.1118 Section 180.1118 Protection of... polyhedrosis virus; exemption from the requirement of a tolerance. An exemption from the requirement of a tolerance is established for the microbial pest control agent Spodoptera exigua nuclear polyhedrosis...

... virus; exemption from the requirement of a tolerance. 180.1118 Section 180.1118 Protection of... polyhedrosis virus; exemption from the requirement of a tolerance. An exemption from the requirement of a tolerance is established for the microbial pest control agent Spodoptera exigua nuclear polyhedrosis...

... virus; exemption from the requirement of a tolerance. 180.1118 Section 180.1118 Protection of... polyhedrosis virus; exemption from the requirement of a tolerance. An exemption from the requirement of a tolerance is established for the microbial pest control agent Spodoptera exigua nuclear polyhedrosis...

Fall armyworm larvae (FAW), Spodoptera frugiperda (J. E. Smith) were collected from whorl-stage cornfields, between the V2 and V4 stages, in 22 localities of Central, Chiapas, México, called "La Frailesca" during late June 2009 to determine the occurrence of native entomopathogens and parasitic nema...

Spodoptera litura multicapsid nucleopolyhedrovirus (SpltMNPV) odv-e25 is 684 bp long, potentially encoding 227 amino acids with a predicted molecular weight of 24.9 kDa. Homology analysis indicated that SpltMNPV ODV-E25 has 35-65% amino acid identity with that of other known baculoviruses. RT-PCR results revealed that the odv-e25 is transcribed actively at the late stage of infection and the mRNA start site was mapped within a consensus baculovirus late promoter sequence (TTAAG). Western blot analysis of odv-e25 expression with an antiserum made against 6 x His tagged ODV-E25 expressed in Escherichia coli indicated that it was present as a doublet of approximately 27 kDa from 24 h through 96 h in SpltMNPV-infected Spli-221 cells. Similar results were seen on Western blots of Spodoptera exigua (Se)MNPV-infected Se301 cells. Immunofluorescence analysis showed that ODV-E25 was predominantly present in the cytoplasm of SpltMNPV-infected cells and localized to the envelopes of occlusion-derived virus.

A recent shift in managing insect resistance to genetically engineered (GE) maize consists of mixing non-GE seed with GE seed known as “refuge in a bag”, which increases the likelihood of predators encountering both prey fed Bt and prey fed non-Bt maize. We therefore conducted laboratory choice-test feeding studies to determine if a predator, Harmonia axyridis, shows any preference between prey fed Bt and non-Bt maize leaves. The prey species was Spodoptera frugiperda, which were fed Bt maize (MON-810), expressing the single Cry1Ab protein, or non-Bt maize. The predators were third instar larvae and female adults of H. axyridis. Individual predators were offered Bt and non-Bt fed prey larvae that had fed for 24, 48 or 72 h. Ten and 15 larvae of each prey type were offered to third instar and adult predators, respectively. Observations of arenas were conducted at 1, 2, 3, 6, 15 and 24 h after the start of the experiment to determine the number and type of prey eaten by each individual predator. Prey larvae that fed on non-Bt leaves were significantly larger than larvae fed Bt leaves. Both predator stages had eaten nearly all the prey by the end of the experiment. However, in all combinations of predator stage and prey age, the number of each prey type consumed did not differ significantly. ELISA measurements confirmed the presence of Cry1Ab in leaf tissue (23–33 µg/g dry weight) and S. frugiperda (2.1–2.2 µg/g), while mean concentrations in H. axyridis were very low (0.01–0.2 µg/g). These results confirm the predatory status of H. axyridis on S. frugiperda and that both H. axyridis adults and larvae show no preference between prey types. The lack of preference between Bt-fed and non-Bt-fed prey should act in favor of insect resistance management strategies using mixtures of GE and non-GE maize seed. PMID:23024772

A recent shift in managing insect resistance to genetically engineered (GE) maize consists of mixing non-GE seed with GE seed known as "refuge in a bag", which increases the likelihood of predators encountering both prey fed Bt and prey fed non-Bt maize. We therefore conducted laboratory choice-test feeding studies to determine if a predator, Harmonia axyridis, shows any preference between prey fed Bt and non-Bt maize leaves. The prey species was Spodoptera frugiperda, which were fed Bt maize (MON-810), expressing the single Cry1Ab protein, or non-Bt maize. The predators were third instar larvae and female adults of H. axyridis. Individual predators were offered Bt and non-Bt fed prey larvae that had fed for 24, 48 or 72 h. Ten and 15 larvae of each prey type were offered to third instar and adult predators, respectively. Observations of arenas were conducted at 1, 2, 3, 6, 15 and 24 h after the start of the experiment to determine the number and type of prey eaten by each individual predator. Prey larvae that fed on non-Bt leaves were significantly larger than larvae fed Bt leaves. Both predator stages had eaten nearly all the prey by the end of the experiment. However, in all combinations of predator stage and prey age, the number of each prey type consumed did not differ significantly. ELISA measurements confirmed the presence of Cry1Ab in leaf tissue (23-33 µg/g dry weight) and S. frugiperda (2.1-2.2 µg/g), while mean concentrations in H. axyridis were very low (0.01-0.2 µg/g). These results confirm the predatory status of H. axyridis on S. frugiperda and that both H. axyridis adults and larvae show no preference between prey types. The lack of preference between Bt-fed and non-Bt-fed prey should act in favor of insect resistance management strategies using mixtures of GE and non-GE maize seed.

The entomopathogenic fungus Beauveria bassiana acts slowly on insect pests through cuticle infection. Vegetative insecticidal proteins (Vip3A) of Bacillus thuringiensis kill lepidopteran pests rapidly, via per os infection, but their use for pest control is restricted to integration into transgenic plants. A transgenic B. bassiana strain (BbV28) expressing Vip3Aa1 (a Vip3A toxin) was thus created to infect the larvae of the oriental leafworm moth Spodoptera litura through conidial ingestion and cuticle adhesion. Vip3Aa1 (∼88 kDa) was highly expressed in the conidial cytoplasm of BbV28 and was detected as a digested form (∼62 kDa) in the larval midgut 18 and 36 h after conidial ingestion. The median lethal concentration (LC50) of BbV28 against the second-instar larvae feeding on cabbage leaves sprayed with conidial suspensions was 26.2-fold lower than that of the wild-type strain on day 3 and 1.1-fold lower on day 7. The same sprays applied to both larvae and leaves for their feeding reduced the LC50 of the transformant 17.2- and 1.3-fold on days 3 and 7, respectively. Median lethal times (LT50s) of BbV28 were shortened by 23 to 35%, declining with conidial concentrations. The larvae infected by ingestion of BbV28 conidia showed typical symptoms of Vip3A action, i.e., shrinkage and palsy. However, neither LC50 nor LT50 trends differed between BbV28 and its parental strain if the infection occurred through the cuticle only. Our findings indicate that fungal conidia can be used as vectors for spreading the highly insecticidal Vip3A protein for control of foliage feeders such as S. litura. PMID:20495052

The entomopathogenic fungus Beauveria bassiana acts slowly on insect pests through cuticle infection. Vegetative insecticidal proteins (Vip3A) of Bacillus thuringiensis kill lepidopteran pests rapidly, via per os infection, but their use for pest control is restricted to integration into transgenic plants. A transgenic B. bassiana strain (BbV28) expressing Vip3Aa1 (a Vip3A toxin) was thus created to infect the larvae of the oriental leafworm moth Spodoptera litura through conidial ingestion and cuticle adhesion. Vip3Aa1 ( approximately 88 kDa) was highly expressed in the conidial cytoplasm of BbV28 and was detected as a digested form ( approximately 62 kDa) in the larval midgut 18 and 36 h after conidial ingestion. The median lethal concentration (LC(50)) of BbV28 against the second-instar larvae feeding on cabbage leaves sprayed with conidial suspensions was 26.2-fold lower than that of the wild-type strain on day 3 and 1.1-fold lower on day 7. The same sprays applied to both larvae and leaves for their feeding reduced the LC(50) of the transformant 17.2- and 1.3-fold on days 3 and 7, respectively. Median lethal times (LT(50)s) of BbV28 were shortened by 23 to 35%, declining with conidial concentrations. The larvae infected by ingestion of BbV28 conidia showed typical symptoms of Vip3A action, i.e., shrinkage and palsy. However, neither LC(50) nor LT(50) trends differed between BbV28 and its parental strain if the infection occurred through the cuticle only. Our findings indicate that fungal conidia can be used as vectors for spreading the highly insecticidal Vip3A protein for control of foliage feeders such as S. litura.

The polyphagous pests belonging to the genus Spodoptera are considered to be among the most important causes of damage and are widely distributed throughout the Americas'. Due to the extensive use of genetically modified plants containing Bacillus thuringiensis genes that code for insecticidal proteins, resistant insects may arise. To prevent the development of resistance, pyramided plants, which express multiple insecticidal proteins that act through distinct mode of actions, can be used. This study analyzed the mechanisms of action for the proteins Cry1Ia10 and Vip3Aa on neonatal Spodoptera frugiperda, Spodoptera albula, Spodoptera eridania and Spodoptera cosmioides larvae. The interactions of these toxins with receptors on the intestinal epithelial membrane were also analyzed by binding biotinylated toxins to brush border membrane vesicles (BBMVs) from the intestines of these insects. A putative receptor of approximately 65 kDa was found by ligand blotting in all of these species. In vitro competition assays using biotinylated proteins have indicated that Vip3Aa and Cry1Ia10 do not compete for the same receptor for S. frugiperda, S. albula and S. cosmioides and that Vip3Aa was more efficient than Cry1Ia10 when tested individually, by bioassays. A synergistic effect of the toxins in S. frugiperda, S. albula and S. cosmioides was observed when they were combined. However, in S. eridania, Cry1Ia10 and Vip3Aa might compete for the same receptor and through bioassays Cry1Ia10 was more efficient than Vip3Aa and showed an antagonistic effect when the proteins were combined. These results suggest that using these genes to develop pyramided plants may not prove effective in preventing the development of resistance in S. eridiana.

Larvae of Culicoides sonorensis a blood feeding ceratopogonid are aquatic. In their natural habitat they are exposed to numerous predators. Predator avoidance behavior has not been studied in most Culicoides spp. Based on laboratory data the larvae of Culicoides sonorensis behave differently when cn...

The incubation period of Spodoptera exigua (Hübner) was not influenced by the host plant, whereas larval development time and pupal period were affected. Larval development time was longest on shallot and lady's finger, followed by cabbage and long bean. Larvae did not develop beyond the first instar when fed on chilli. The pupal period was longer on lady's finger than on cabbage, shallot and long bean. Overall, adult longevity was not influenced by the host plant but there was a difference between female and male longevity among the host plants. Survival of S. exigua was affected by the host plant at the larval stage. The number of larval instars varied between 5 and 8 within and between the studied host plants. Long bean was found to be the most suitable host plant and provide the best food quality for S. exigua compared to the other host plants, as it allowed faster development, fewer larval instars and a higher survival rate.

The complete mitochondrial genome (mitogenome) of Spodoptera litura (Lepidoptera: Noctuidae) was determined to be 15,374 bp (GenBank accession No. KF543065), including 13 protein-coding genes (PCGs), two rRNA genes, 22 tRNA genes and an A + T-rich region. It has the typical gene organization and order of mitogenomes from lepidopteran insects. The AT skew of this mitogenome was slightly positive and the nucleotide composition was also biased toward A + T nucleotides (81.03%). All PCGs were initiated by ATN codons, except for cytochrome c oxidase subunit 1 (cox1) gene which was initiated by CGA. Four of the 13 PCGs harbor the incomplete termination codon by T. All the tRNA genes displayed a typical clover-leaf structure of mitochondrial tRNA, with the exception of trnS1 (AGN). The A + T-rich region of the mitogenome was 326 bp in length.

The vegetative insecticidal proteins (Vip) secreted by many Bacillus thuringiensis strains during their vegetative growth stage are regarded as second generation insecticidal proteins, as they share no sequence or structural homology with known crystal insecticidal proteins (Cry) and have a broad insecticidal spectrum. Compared with insecticidal crystal proteins (ICPs), the insecticidal mechanisms of Vips have been little studied. Here we investigated the mechanism responsible for Vip3Aa toxicity in cultured insect cells. Using, flow cytometry analyzes, TUNEL staining and DNA fragmentation assays, we show that Vip3Aa can induce apoptosis in Spodoptera frugiperda (Sf9) cells and cause cells to arrest at the G2/M phase. We also show that Vip3Aa can disrupt mitochondrial membrane potential (ΔΨm), leading to the activation of Sf-caspase-1, suggesting that a mitochondrial mediated and caspase dependent pathway may be involved in Vip3Aa-induced apoptosis in Sf9 cells.

The seeds of Phaseolus lathyroides were extracted with hexane, dichloromethane, ethyl acetate and methanol, respectively by Soxhlet apparatus. Each crude extract was examined for toxicity against the second instars of Spodoptera litura using a topical application method under laboratory conditions. The ethyl acetate extract showed the most effective mortality (LD₅₀ = 11,964 and 9,169 ppm after treated at 24 and 48 hours, respectively). Furthermore, in vivo enzyme based experiments revealed that acetylcholinesterase activity of survived S. litura (24 hours post-treatment) was increased by 12% compared to control experiments. Our result showed the possibility to develop alternative strategies by using extract from the seeds of Phaseolus lathyroides for the control of S. litura.

Male moths can finely discriminate the sex pheromone emitted by conspecific females from similar compounds. Pheromone receptors, expressed on the dendritic membrane of sensory neurons housed in the long trichoid sensilla of antennae, are thought to be associated with the pheromone reception. In this study, we identified and functionally characterized 4 pheromone receptors from the antennae of Spodoptera litura (Lepidoptera: Noctuidae). A tissue distribution analysis showed that the expression of the 4 SlituPRs was restricted to antennae. In addition, SlituOR6 and SlituOR13 were specifically expressed in male antennae whereas SlituOR11 and SlituOR16 were male-biased. Functional investigation by heterologous expression in Xenopus oocytes revealed that SlituOR6 was specifically tuned to the second major pheromone component, Z9,E12-14:OAc, SlituOR13 was equally tuned to Z9,E12-14:OAc and Z9-14:OAc, with a small response to the major pheromone component Z9,E11-14:OAc, SlituOR16 significantly responded to the behavioral antagonist Z9-14:OH, whereas SlituOR11 did not show response to any of the pheromone compounds tested in this study. Our results provide molecular data to better understand the mechanisms of sex pheromone detection in the moth S. litura and bring clues to investigate the evolution of the sexual communication channel in closely related species through comparison with previously reported pheromone receptors in other Spodoptera species.

In this study, we examined the prevalence, pathology, and epidemiology of tumors in free-ranging island foxes occurring on three islands in the California Channel Islands, USA. We found a remarkably high prevalence of ceruminous gland tumors in endangered foxes (Urocyon littoralis catalinae) occurring on Santa Catalina Island (SCA)—48.9% of the dead foxes examined from 2001–2008 had tumors in their ears, and tumors were found in 52.2% of randomly-selected mature (≥ 4 years) foxes captured in 2007–2008, representing one of the highest prevalences of tumors ever documented in a wildlife population. In contrast, no tumors were detected in foxes from San Nicolas Island or San Clemente Island, although ear mites (Otodectes cynotis), a predisposing factor for ceruminous gland tumors in dogs and cats, were highly prevalent on all three islands. On SCA, otitis externa secondary to ear mite infection was highly correlated with ceruminous gland hyperplasia (CGH), and tumors were significantly associated with the severity of CGH, ceruminous gland dysplasia, and age group (older foxes). We propose a conceptual model for the formation of ceruminous gland tumors in foxes on SCA that is based on persistent, ubiquitous infection with ear mites, and an innate, over exuberant inflammatory and hyperplastic response of SCA foxes to these mites. Foxes on SCA are now opportunistically treated with acaricides in an attempt to reduce mite infections and the morbidity and mortality associated with this highly prevalent tumor. PMID:26618759

In this study, we examined the prevalence, pathology, and epidemiology of tumors in free-ranging island foxes occurring on three islands in the California Channel Islands, USA. We found a remarkably high prevalence of ceruminous gland tumors in endangered foxes (Urocyon littoralis catalinae) occurring on Santa Catalina Island (SCA)--48.9% of the dead foxes examined from 2001-2008 had tumors in their ears, and tumors were found in 52.2% of randomly-selected mature (≥ 4 years) foxes captured in 2007-2008, representing one of the highest prevalences of tumors ever documented in a wildlife population. In contrast, no tumors were detected in foxes from San Nicolas Island or San Clemente Island, although ear mites (Otodectes cynotis), a predisposing factor for ceruminous gland tumors in dogs and cats, were highly prevalent on all three islands. On SCA, otitis externa secondary to ear mite infection was highly correlated with ceruminous gland hyperplasia (CGH), and tumors were significantly associated with the severity of CGH, ceruminous gland dysplasia, and age group (older foxes). We propose a conceptual model for the formation of ceruminous gland tumors in foxes on SCA that is based on persistent, ubiquitous infection with ear mites, and an innate, over exuberant inflammatory and hyperplastic response of SCA foxes to these mites. Foxes on SCA are now opportunistically treated with acaricides in an attempt to reduce mite infections and the morbidity and mortality associated with this highly prevalent tumor.

Alkaloids are the most extensively featured compounds of natural anti-tumor herbs, which have attracted much attention in pharmaceutical research. In our previous studies, a mixture of major three alkaloid components (5, 6-dihydrobicolorine, 7-deoxy-trans-dihydronarciclasine, littoraline) from Hymenocallis littoralis were extracted, analyzed and designated as AHL. In this paper, AHL extracts were added to human liver hepatocellular cells HepG-2, human gastric cancer cell SGC-7901, human breast adenocarcinoma cell MCF-7 and human umbilical vein endothelial cell EVC-304, to screen one or more AHL-sensitive tumor cell. Among these cells, HepG-2 was the most sensitive to AHL treatment, a very low dose (0.8μg/ml) significantly inhibiting proliferation . The non- tumor cell EVC-304, however, was not apparently affected. Effect of AHL on HepG-2 cells was then explored. We found that the AHL could cause HepG-2 cycle arrest at G2/M checkpoint, induce apoptosis, and interrupt polymerization of microtubules. In addition, expression of two cell cycle-regulated proteins, CyclinB1 and CDK1, was up-regulated upon AHL treatment. Up-regulation of the Fas, Fas ligand, Caspase-8 and Caspase-3 was observed as well, which might imply roles for the Fas/FsaL signaling pathway in the AHL-induced apoptosis of HepG-2 cells.

Oomycete diseases in seaweeds are probably widespread and of significant ecological and economic impact, but overall still poorly understood. This study investigates the organisation of the cytoskeleton during infection of three brown algal species, Pylaiella littoralis, Ectocarpus siliculosus, and Ectocarpus crouaniorum, by the basal marine oomycete Eurychasma dicksonii. Immunofluorescence staining of tubulin revealed how the development of this intracellular biotrophic pathogen impacts on microtubule (MT) organisation of its algal host. The host MT cytoskeleton remains normal and organised by the centrosome until very late stages of the infection. Additionally, the organisation of the parasite's cytoskeleton was examined. During mitosis of the E. dicksonii nucleus the MT focal point (microtubule organisation centre, MTOC, putative centrosome) duplicates and each daughter MTOC migrates to opposite poles of the nucleus. This similarity in MT organisation between the host and pathogen reflects the relatively close phylogenetic relationship between oomycetes and brown algae. Moreover, actin labelling with rhodamine-phalloidin in E. dicksonii revealed typical images of actin dots connected by fine actin filament bundles in the cortical cytoplasm. The functional and phylogenetic implications of our observations are discussed.

Background Transgenic crops expressing Bt toxins have substantial benefits for growers in terms of reduced synthetic insecticide inputs, area-wide pest management and yield. This valuable technology depends upon delaying the evolution of resistance. The ‘high dose/refuge strategy’, in which a refuge of non-Bt plants is planted in close proximity to the Bt crop, is the foundation of most existing resistance management. Most theoretical analyses of the high dose/refuge strategy assume random oviposition across refugia and Bt crops. Results In this study we examined oviposition and survival of Spodoptera frugiperda across conventional and Bt maize and explored the impact of oviposition behavior on the evolution of resistance in simulation models. Over six growing seasons oviposition rates per plant were higher in Bt crops than in refugia. The Cry1F Bt maize variety retained largely undamaged leaves, and oviposition preference was correlated with the level of feeding damage in the refuge. In simulation models, damage-avoiding oviposition accelerated the evolution of resistance and either led to requirements for larger refugia or undermined resistance management altogether. Since larval densities affected oviposition preferences, pest population dynamics affected resistance evolution: larger refugia were weakly beneficial for resistance management if they increased pest population sizes and the concomitant degree of leaf damage. Conclusions Damaged host plants have reduced attractiveness to many insect pests, and crops expressing Bt toxins are generally less damaged than conventional counterparts. Resistance management strategies should take account of this behavior, as it has the potential to undermine the effectiveness of existing practice, especially in the tropics where many pests are polyvoltinous. Efforts to bring down total pest population sizes and/or increase the attractiveness of damaged conventional plants will have substantial benefits for slowing the

Brazil ranked second only to the United States in hectares planted to genetically modified crops in 2013. Recently corn producers in the Cerrado region reported that the control of Spodoptera frugiperda with Bt corn expressing Cry1Fa has decreased, forcing them to use chemicals to reduce the damage caused by this insect pest. A colony of S. frugiperda was established from individuals collected in 2013 from Cry1Fa corn plants (SfBt) in Brazil and shown to have at least more than ten-fold higher resistance levels compared with a susceptible colony (Sflab). Laboratory assays on corn leaves showed that in contrast to SfLab population, the SfBt larvae were able to survive by feeding on Cry1Fa corn leaves. The SfBt population was maintained without selection for eight generations and shown to maintain high levels of resistance to Cry1Fa toxin. SfBt showed higher cross-resistance to Cry1Aa than to Cry1Ab or Cry1Ac toxins. As previously reported, Cry1A toxins competed the binding of Cry1Fa to brush border membrane vesicles (BBMV) from SfLab insects, explaining cross-resistance to Cry1A toxins. In contrast Cry2A toxins did not compete Cry1Fa binding to SfLab-BBMV and no cross-resistance to Cry2A was observed, although Cry2A toxins show low toxicity to S. frugiperda. Bioassays with Cry1AbMod and Cry1AcMod show that they are highly active against both the SfLab and the SfBt populations. The bioassay data reported here show that insects collected from Cry1Fa corn in the Cerrado region were resistant to Cry1Fa suggesting that resistance contributed to field failures of Cry1Fa corn to control S. frugiperda.

Chitin synthase (CHS) is an important enzymatic component, which is required for chitin formation in the cuticles and cuticular linings of other tissues in insects. CHSs have been divided into two classes, classes A and B, based on their amino acid sequence similarities and functions. Class A CHS (CHS-A) is specifically expressed in the epidermis and related ectodermal cells such as tracheal cells, while class B CHS (CHS-B) is expressed in gut epithelial cells that produce peritrophic matrices. In this study, we cloned the CHS-A gene from the beet armyworm, Spodoptera exigua (SeCHS-A). The SeCHS-A contains an open reading frame of 4,698 nucleotides, encoding a protein of 1,565 amino acids with a predicted molecular mass of approximately 177.8 kDa. The SeCHS-A mRNA was expressed in all developmental stages and specifically in the epidermis and tracheae tissue by quantitative real-time-PCR analysis. Expression of SeCHS-A gene was suppressed by feeding double-stranded RNA (dsCHS-A, 400 ng/larva) in the third instar larvae of S. exigua. Suppression of the SeCHS-A gene expression significantly increased 35% of mortality on pupation of S. exigua. Also, the third instar larvae fed with dsCHS-A significantly increased susceptibility to entomopathogenic fungi, Beauveria bassiana ANU1 at 3 days after treatment. These results suggest that the SeCHS-A gene plays an important role in development of S. exigua and RNA interference may apply to effective pest control with B. bassiana.

Biological control provides a safer alternative to reduce the population of agricultural pest. Mirabilis jalapa is one of many promising biopesticides which contains chemical substances that have a feeding deterrent property against insects. This biopesticide may not kill insect directly but will weaken their overall physiological condition. In this study, we investigated the immune response of common pestSpodoptera litura after exposure of M. jalapa extract. We also used Bacillus thuringiensis (Bt) delta endotoxin (LC50) on 3 hours after exposure of M. jalapa extract to see the synergism properties of both biopesticide agents. Microscopic observation revealed that at least 5 types of haemocyte were found in S. litura. In control group, plasmatocyte were found at 59.98%, prohaemocyte 20.73%, granullar cell 12.74%, oenocytoid 3.33% and spherule cell 3.20%. These proportion was differ significantly in the treatment group. Exposure to 0.1% and 0.2%(w/v) of M. jalapa extract increased the total number of haemocytes as much as 38.08% and 64.15% respectively. In contrast, exposure to 0.4% and 0.8%(w/v) reduced the number of haemocytes to 37.02% and 51.04% respectively. In term of phagocytic activity, the proportion of phagocytosing cells were 47.62% in control group, and in 0.1% and 0.2% (w/v) M. jalapa treatment group the proportion decreased to 28% and 26.88% respectively. In the concentration of 0.4% and 0.8%, phagocytic activity did not occur. Addition of biological agents Bt (LC50 concentration) to see mortality 3 hours after M. jalapa application did not show significant differences. S. litura mortality rate were found only 50%; this suggests that the combination of M. jalapa and Bt biopesticides in 3-hour intervals within 24 hours showed no increase in mortality.

Host-pathogen interactions result in complex relationship, many aspects of which are not completely understood. Vip proteins, which are Bacillus thuringensis (Bt) insecticidal toxins produced during the vegetative stage, are selectively effective against specific insect pests. This new group of Bt proteins represents an interesting alternative to the classical Bt Cry toxins because current data suggests that they do not share the same mode of action. We have designed and developed a genome-wide microarray for the beet armyworm Spodoptera exigua, a serious lepidopteran pest of many agricultural crops, and used it to better understand how lepidopteran larvae respond to the treatment with the insecticidal protein Vip3Aa. With this approach, the goal of our study was to evaluate the changes in gene expression levels caused by treatment with sublethal doses of Vip3Aa (causing 99% growth inhibition) at 8 and 24 h after feeding. Results indicated that the toxin provoked a wide transcriptional response, with 19% of the microarray unigenes responding significantly to treatment. The number of up- and down-regulated unigenes was very similar. The number of genes whose expression was regulated at 8 h was similar to the number of genes whose expression was regulated after 24 h of treatment. The up-regulated sequences were enriched for genes involved in innate immune response and in pathogen response such as antimicrobial peptides (AMPs) and repat genes. The down-regulated sequences were mainly unigenes with homology to genes involved in metabolism. Genes related to the mode of action of Bt Cry proteins were found, in general, to be slightly overexpressed. The present study is the first genome-wide analysis of the response of lepidopteran insects to Vip3Aa intoxication. An insight into the molecular mechanisms and components related to Vip intoxication will allow designing of more effective management strategies for pest control. PMID:24312604

Host-pathogen interactions result in complex relationship, many aspects of which are not completely understood. Vip proteins, which are Bacillus thuringensis (Bt) insecticidal toxins produced during the vegetative stage, are selectively effective against specific insect pests. This new group of Bt proteins represents an interesting alternative to the classical Bt Cry toxins because current data suggests that they do not share the same mode of action. We have designed and developed a genome-wide microarray for the beet armyworm Spodoptera exigua, a serious lepidopteran pest of many agricultural crops, and used it to better understand how lepidopteran larvae respond to the treatment with the insecticidal protein Vip3Aa. With this approach, the goal of our study was to evaluate the changes in gene expression levels caused by treatment with sublethal doses of Vip3Aa (causing 99% growth inhibition) at 8 and 24 h after feeding. Results indicated that the toxin provoked a wide transcriptional response, with 19% of the microarray unigenes responding significantly to treatment. The number of up- and down-regulated unigenes was very similar. The number of genes whose expression was regulated at 8 h was similar to the number of genes whose expression was regulated after 24 h of treatment. The up-regulated sequences were enriched for genes involved in innate immune response and in pathogen response such as antimicrobial peptides (AMPs) and repat genes. The down-regulated sequences were mainly unigenes with homology to genes involved in metabolism. Genes related to the mode of action of Bt Cry proteins were found, in general, to be slightly overexpressed. The present study is the first genome-wide analysis of the response of lepidopteran insects to Vip3Aa intoxication. An insight into the molecular mechanisms and components related to Vip intoxication will allow designing of more effective management strategies for pest control.

The Cry proteins produced by Bacillus thuringiensis (Bt) are the most widely used biopesticides effective against a range of crop pests and disease vectors. Like chemical pesticides, development of resistance is the primary threat to the long-term efficacy of Bt toxins. Recently discovered cadherin-based Bt Cry synergists showed the potential to augment resistance management by improving efficacy of Cry toxins. However, the mode of action of Bt Cry synergists is thus far unclear. Here we elucidate the mechanism of cadherin-based Cry toxin synergism utilizing two cadherin peptides, Spodoptera frugiperda Cad (SfCad) and Manduca sexta Cad (MsCad), which differentially enhance Cry1Fa toxicity to Spodoptera frugiperda neonates. We show that differential SfCad- and MsCad-mediated protection of Cry1Fa toxin in the Spodoptera frugiperda midgut correlates with differential Cry1Fa toxicity enhancement. Both peptides exhibited high affinity for Cry1Fa toxin and an increased rate of Cry1Fa-induced pore formation in S. frugiperda. However, only SfCad bound the S. frugiperda brush border membrane vesicle and more effectively prolonged the stability of Cry1Fa toxin in the gut, explaining higher Cry1Fa enhancement by this peptide. This study shows that cadherin fragments may enhance B. thuringiensis toxicity by at least two different mechanisms or a combination thereof: (i) protection of Cry toxin from protease degradation in the insect midgut and (ii) enhancement of pore-forming ability of Cry toxin. PMID:22081566

Flea beetles of alligator weed, Alternanthera philoxeroides (Martius) Grisebach (Amaranthaceae), were collected in Argentina, Uruguay, Paraguay, and Brazil. Species in the genera Disonycha Chevrolat, Agasicles Jacoby, Systena Chevrolat and Phenrica Bechyné were frequently found on this weed. Phenrica littoralis (Bechyn6) was the most abundant within this genus. The male is described and the holotype female is redescribed adding new diagnostic characters of the mouthparts, hind wings, metendosternite, and male and female genitalia. Larva and pupa are described and illustrated for the first time providing data for future phylogenetic studies in the subtribe Disonychina.

Caecilians are limbless amphibians that have evolved distinct cranial and postcranial specializations associated with a burrowing lifestyle. Observations on feeding behavior are rare and restricted to above-ground feeding in laboratory conditions. Here we report data on feeding in tunnels using both external video and X-ray recordings of caecilians feeding on invertebrate prey. Our data show feeding kinematics similar to those previously reported, including the pronounced neck bending observed during above-ground feeding. Our data illustrate, however, that caecilians may be much faster than previously suspected, with lunge speeds of up to 7 cm sec(-1). Although gape cycles are often slow (0.67 ± 0.29 sec), rapid jaw closure is observed during prey capture, with cycle times and jaw movement velocities similar to those observed in other terrestrial tetrapods. Finally, our data suggest that gape angles may be large (64.8 ± 18°) and that gape profiles are variable, often lacking distinct slow and fast opening and closing phases. These data illustrate the importance of recording naturalistic feeding behavior and shed light on how these animals are capable of capturing and processing prey in constrained underground environments. Additional data on species with divergent cranial morphologies would be needed to better understand the co-evolution between feeding, burrowing, and cranial design in caecilians.

The evolutionary mechanisms generating the tremendous biodiversity of islands have long fascinated evolutionary biologists. Genetic drift and divergent selection are predicted to be strong on islands and both could drive population divergence and speciation. Alternatively, strong genetic drift may preclude adaptation. We conducted a genomic analysis to test the roles of genetic drift and divergent selection in causing genetic differentiation among populations of the island fox (Urocyon littoralis). This species consists of six subspecies, each of which occupies a different California Channel Island. Analysis of 5293 SNP loci generated using Restriction-site Associated DNA (RAD) sequencing found support for genetic drift as the dominant evolutionary mechanism driving population divergence among island fox populations. In particular, populations had exceptionally low genetic variation, small Ne (range = 2.1-89.7; median = 19.4), and significant genetic signatures of bottlenecks. Moreover, islands with the lowest genetic variation (and, by inference, the strongest historical genetic drift) were most genetically differentiated from mainland grey foxes, and vice versa, indicating genetic drift drives genome-wide divergence. Nonetheless, outlier tests identified 3.6-6.6% of loci as high FST outliers, suggesting that despite strong genetic drift, divergent selection contributes to population divergence. Patterns of similarity among populations based on high FST outliers mirrored patterns based on morphology, providing additional evidence that outliers reflect adaptive divergence. Extremely low genetic variation and small Ne in some island fox populations, particularly on San Nicolas Island, suggest that they may be vulnerable to fixation of deleterious alleles, decreased fitness and reduced adaptive potential.

We report here the isolation and functional analysis of AlTMP1 gene encoding a member of the PMP3 protein family. In Aeluropus littoralis, AlTMP1 is highly induced by abscisic acid (ABA), cold, salt, and osmotic stresses. Transgenic tobacco expressing AlTMP1 exhibited enhanced tolerance to salt, osmotic, H₂O₂, heat and freezing stresses at the seedling stage. Under greenhouse conditions, the transgenic plants showed a higher level of tolerance to drought than to salinity. Noteworthy, AlTMP1 plants yielded two- and five-fold more seeds than non-transgenic plants (NT) under salt and drought stresses, respectively. The leaves of AlTMP1 plants accumulated lower Na⁺ but higher K⁺ and Ca(2+) than those of NT plants. Tolerance to osmotic and salt stresses was associated with higher membrane stability, low electrolyte leakage, and improved water status. Finally, accumulation of AlTMP1 in tobacco altered the regulation of some stress-related genes in either a positive (NHX1, CAT1, APX1, and DREB1A) or negative (HKT1 and KT1) manner that could be related to the observed tolerance. These results suggest that AlTMP1 confers stress tolerance in tobacco through maintenance of ion homeostasis, increased membrane integrity, and water status. The observed tolerance may be due to a direct or indirect effect of AlTMP1 on the expression of stress-related genes which could stimulate an adaptive potential not present in NT plants.

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Spodoptera frugiperda (Lepidoptera: Noctuidae) is a major pest in maize crops in Colombia, and affects several regions in America. A granulovirus isolated from S. frugiperda (SfGV VG008) has potential as an enhancer of insecticidal activity of previously described nucleopolyhedrovirus from the same insect species (SfMNPV). The SfGV VG008 genome was sequenced and analyzed showing circular double stranded DNA of 140,913 bp encoding 146 putative ORFs that include 37 Baculoviridae core genes, 88 shared with betabaculoviruses, two shared only with betabaculoviruses from Noctuide insects, two shared with alphabaculoviruses, three copies of own genes (paralogs) and the other 14 corresponding to unique genes without representation in the other baculovirus species. Particularly, the genome encodes for important virulence factors such as 4 chitinases and 2 enhancins. The sequence analysis revealed the existence of eight homologous regions (hrs) and also suggests processes of gene acquisition by horizontal transfer including the SfGV VG008 ORFs 046/047 (paralogs), 059, 089 and 099. The bioinformatics evidence indicates that the genome donors of mentioned genes could be alpha- and/or betabaculovirus species. The previous reported ability of SfGV VG008 to naturally co-infect the same host with other virus show a possible mechanism to capture genes and thus improve its fitness.

Reproductive toxicity of Zn to insects was investigated in this study. By exposing phytophagous insect Spodoptera litura Fabricius to Zn in artificial diets of larvae, we investigated the effects of Zn on reproduction at ecological and molecular levels. A significantly shorter period of laying eggs was observed in S. litura exposed to 300-750mg Zn/kg. The oviposition rate, fecundity and hatchability of female adults treated with 750mg Zn/kg were significantly lower than those of the controls (31.43%, 20.95% and 52%, respectively, compared to the control). The Zn accumulation and vitellin (Vn) content in eggs were tested by inductively coupled plasma-atomic emission spectrometry and Bradford combining Western-blot, respectively. The results showed that Zn accumulated in the eggs, which has affected the weight and Vn content of eggs with significant negative correlations. The down-regulated expression levels of vitellogenin (Vg) mRNA were detected by real-time polymerase chain reaction (RT-PCR): the relative quantity of Vg mRNA was less than half of the controls at higher than 450mg Zn/kg wet weight. These results indicated that excess Zn made expression of Vg gene down-regulated and caused poor accumulation of egg yolk, which led to a reduction in egg numbers and failure of eggs to hatch.

The beet armyworm Spodoptera exigua (Lepidoptera: Noctuidae) is difficult to control using chemical insecticides because of the development of insecticide resistance. Several pest control agents are used to control the beet armyworm. Entomopathogenic fungi are one of the candidates for eco-friendly pest control instead of chemical control agents. In this study, among various entomopathogenic fungal strains isolated from soil two isolates were selected as high virulence pathogens against larva of beet armyworm. Control efficacy of fungal conidia was influenced by conidia concentration, temperature, and relative humidity (RH). The isolates Metarhizium anisopliae FT83 showed 100% cumulative mortality against second instar larvae of S. exigua 3 days after treatment at 1 × 107 conidia/mL and Paecilomyces fumosoroseus FG340 caused 100% mortality 6 days after treatment at 1 × 104 conidia/mL. Both M. anisopliae FT83 and P. fumosoroseus FG340 effectively controlled the moth at 20~30℃. M. anisopliae FT83 was significantly affected mortality by RH: mortality was 86.7% at 85% RH and 13.4% at 45% RH. P. fumosoroseus FG340 showed high mortality as 90% at 45% RH and 100% at 75% RH 6 days after conidia treatments. These results suggest that P. fumosoroseus FG340 and M. anisopliae FT83 have high potential to develop as a biocontrol agent against the beet armyworm. PMID:25606011

Two forms of acetylcholinesterase were identified in field populations of the beet armyworm, Spodoptera exigua (Hübner), collected from cotton in San Joaquin Valley, CA. Strains (BESS and BKRR) homogeneous for each variant were isolated and their relative susceptibilities to methomyl, chlorpyrifos, and chlorpyrifos-oxon assessed by topical application bioassay. In comparisons with a laboratory susceptible strain (DOW), BKRR and BESS expressed 68-fold and sevenfold resistance, respectively, to the carbamate methomyl. Neither strain was cross-resistant to chlorpyrifos or its oxygen analog (chlorpyrifos-oxon). In biochemical studies, the BKRR AChE enzyme was approximately 30-fold and sixfold more insensitive to methomyl and chlorpyrifos-oxon, respectively, compared with the DOW enzyme. The correlation between the toxicological and biochemical studies provides strong evidence that target-site insensitivity is an important mechanism of resistance to methomyl. The lack of significant cross-resistance to chlorpyrifos suggests that the insensitive AChE in these field populations was selected by methomyl alone and not by the organophosphate.

Digestive enzymatic activity in three geographic strains (Miandiab, Kalposh and Moghan regions) of Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae) reared on different sugar beet cultivars (Dorothea, Rozier, Persia and Perimer) was studied under laboratory conditions (25 ± 1 °C, 65 ± 5% RH, and a photo period of 16:8 (L:D) h photoperiod). The results of this study demonstrated that digestive protease and amylase activity of S. exigua larvae was affected by both geographic origin of the pest and host plant cultivar. Three strains reared on the same sugar beet cultivars demonstrated different levels of proteolytic and amylolytic activities in fourth and fifth instars. The highest proteolytic and amylolytic activity, in most cases, was observed in larvae collected from Kalposh region. Among different sugar beet cultivars, the highest protease activity in three strains was observed on cultivars Rozier and Perimer. Nevertheless, the highest amylase activity was seen on cultivar Dorothea, and the lowest activity was seen on cultivar Rozier. This study suggested that variations in digestive enzymatic activity of three geographic strains of S. exigua might be attributed to local adaptation with their local host plant and environmental conditions inherent by larvae. PMID:28069730

Background Baculovirus comprise the largest group of insect viruses most studied worldwide, mainly because they efficiently kill agricutural insect pests. In this study, two recombinant baculoviruses containing the ScathL gene from Sarcophaga peregrina (vSynScathL), and the Keratinase gene from the fungus Aspergillus fumigatus (vSynKerat), were constructed. and their insecticidal properties analysed against Spodoptera frugiperda larvae. Results Bioassays of third-instar and neonate S. frugiperda larvae with vSynScathL and vSynKerat showed a decrease in the time needed to kill the infected insects when compared to the wild type virus. We have also shown that both recombinants were able to increase phenoloxidase activity in the hemolymph of S. frugiperda larvae. The expression of proteases in infected larvae resulted in destruction of internal tissues late in infection, which could be the reason for the increased viral speed of kill. Conclusions Baculoviruses and their recombinant forms constitute viable alternatives to chemical insecticides. Recombinant baculoviruses containing protease genes can be added to the list of engineered baculoviruses with great potential to be used in integrated pest management programs. PMID:20587066

If the carbon dioxide (CO2) concentration in the atmosphere changes in the future, as predicted, it could influence crops and insect pests. The growth and development of the tobacco caterpillar, Spodoptera litura (Fabricius) (Noctuidae: Lepidoptera), reared on peanut (Arachis hypogea L.) foliage grown under elevated CO2 (550 ppm and 700 ppm) concentrations in open top chambers at Central Research Institute for Dryland Agriculture, Hyderabad, India, were examined in this study. Significantly lower leaf nitrogen, higher carbon, higher relative proportion of carbon to nitrogen and higher polyphenols content expressed in terms of tannic acid equivalents were observed in the peanut foliage grown under elevated CO2 levels. Substantial influence of elevated CO2 on S. litura was noticed, such as longer larval duration, higher larval weights, and increased consumption of peanut foliage by S. litura larvae under elevated CO2 compared with ambient CO2. Relative consumption rate was significantly higher for S. litura larva fed plants grown at 550 and 700 ppm than for larvae fed plants grown at ambient condition. Decreased efficiency of conversion of ingested food, decreased efficiency of conversion of digested food, and decreased relative growth rate of larvae was observed under elevated CO2. The present results indicate that elevated CO2 levels altered the quality of the peanut foliage, resulting in higher consumption, lower digestive efficiency, slower growth, and longer time to pupation (one day more than ambient). PMID:23437971

Although the general pathway of sex pheromone synthesis in moth species has been established, the molecular mechanisms remain poorly understood. The common cutworm Spodoptera litura is an important agricultural pest worldwide and causes huge economic losses annually. The female sex pheromone of S. litura comprises Z9,E11-14:OAc, Z9,E12-14:OAc, Z9-14:OAc, and E11-14:OAc. By sequencing and analyzing the transcriptomic data of the sex pheromone glands, we identified 94 candidate genes related to pheromone biosynthesis (55 genes) or chemoreception (39 genes). Gene expression patterns and phylogenetic analysis revealed that two desaturase genes (SlitDes5 and SlitDes11) and one fatty acyl reductase gene (SlitFAR3) showed pheromone gland (PG) biased or specific expression, and clustered with genes known to be involved in pheromone synthesis in other moth species. Furthermore, 4 chemoreception related genes (SlitOBP6, SlitOBP11, SlitCSP3, and SlitCSP14) also showed higher expression in the PG, and could be additional candidate genes involved in sex pheromone transport. This study provides the first solid background information that should facilitate further elucidation of sex pheromone biosynthesis and transport, and indicates potential targets to disrupt sexual communication in S. litura for a novel pest management strategy. PMID:26445454

Pheromone and plant odorants are important for insect mating, foraging food sources and oviposition. To understand the molecular mechanisms regulating pheromone and odorant signaling, we employed qRT-PCR to study the circadian rhythms of ABP, OBP, PBP, and OR gene expression in the beet armyworm, Spodoptera exigua and their responses after a pre-exposure to sex pheromone compounds or plant volatiles. The neuronal responses of male S. exigua to 20 chemical compounds were recorded at three specific time periods using the electroantennogram. The results showed a circadian rhythm in the expression profiles of some chemosensory genes in the antennae similar to their behavioral rhythm. The expression profiles of OR3, OR6, OR11, OR13, OR16, OR18, Orco, ABP2, OBP1, OBP7, and PBP1, and EAG responses to chemical compounds, as well as their circadian rhythm were significantly affected after exposure to synthetic sex pheromones and plant volatiles. These findings provide the first evidence that the gene expression of chemosensory genes and olfactory sensitivity to sex pheromones are affected by pre-exposing insects to pheromone compounds and plant volatiles. It helps to understand the molecular mechanisms underlying pheromone activity, and the application of sex pheromones and plant volatiles in mating disruption or mass trapping. PMID:26611815

If the carbon dioxide (CO(2)) concentration in the atmosphere changes in the future, as predicted, it could influence crops and insect pests. The growth and development of the tobacco caterpillar, Spodoptera litura (Fabricius) (Noctuidae: Lepidoptera), reared on peanut (Arachis hypogea L.) foliage grown under elevated CO(2) (550 ppm and 700 ppm) concentrations in open top chambers at Central Research Institute for Dryland Agriculture, Hyderabad, India, were examined in this study. Significantly lower leaf nitrogen, higher carbon, higher relative proportion of carbon to nitrogen and higher polyphenols content expressed in terms of tannic acid equivalents were observed in the peanut foliage grown under elevated CO(2) levels. Substantial influence of elevated CO(2) on S. litura was noticed, such as longer larval duration, higher larval weights, and increased consumption of peanut foliage by S. litura larvae under elevated CO(2) compared with ambient CO(2). Relative consumption rate was significantly higher for S. litura larva fed plants grown at 550 and 700 ppm than for larvae fed plants grown at ambient condition. Decreased efficiency of conversion of ingested food, decreased efficiency of conversion of digested food, and decreased relative growth rate of larvae was observed under elevated CO(2). The present results indicate that elevated CO(2) levels altered the quality of the peanut foliage, resulting in higher consumption, lower digestive efficiency, slower growth, and longer time to pupation (one day more than ambient).

The effects of calcium ions and modulators of calcium movement on Bacillus thuringiensis insecticidal protein toxicity were investigated with Sf9 cells (Spodoptera frugiperda, fall armyworm) by a new B. thuringiensis toxicity assay based on measurement of fluorescence of ethidium homodimer, a high-affinity DNA stain. CryIC toxicity was substantially stimulated by extracellular calcium in a dose-dependent way (in the millimolar range), while toxicity enhancement could not be replicated when calcium was replaced by barium. This incremental toxicity was reduced by cobalt and lanthanum ions, two inorganic-calcium transport inhibitors. Methoxyverapamil, a voltage-dependent calcium channel blocker, and nifedipine, an inhibitor of dihydropyridine-sensitive L-type calcium channels, had no effect on CryIC toxin activity, but BAY K 8644, an L-type calcium channel activator, increased CryIC activity at high concentrations of extracellular calcium. While A23187, a calcium ionophore, and TMB-8, an inhibitor of intracellular-calcium mobilization, did not change CryIC-induced mortality, thapsigargin, an inhibitor of calcium uptake in intracellular stores, and more particularly trifluoperazine, which inhibits calcium-calmodulin-dependent processes, increased CryIC-mediated toxicity. The incremental effect of extracellular calcium on CryIC-induced toxicity was consistent with an increased concentration of intracellular calcium. PMID:16535509

The existence of a diverse serine proteinase gene family in lepidopteran insects suggests they play a significant role in the insect adaptation to plant proteinase inhibitors. These proteinases have been shown to be involved in the process of proteolytic digestion in insect larvae. We carried out a selective transcriptome study of midguts from Spodoptera frugiperda larvae fed on a diet supplemented with soybean proteinase inhibitor (SPI). Using subtracted cDNA libraries made of gut-expressed transcripts, a total of 2100 partial sequences were obtained, of those 38% were related to digestive process. Two large and diverse groups of chymotrypsins and trypsins were obtained, and some of these proteinase-encoding genes were further characterized by quantitative RT-PCR. The transcription analyses revealed two groups: one group of genes constitutively expressed in the control larvae that is up regulated by introducing SPI to the diet, and a second group that is absent in the control but is induced by the SPI-rich diet. This observation suggests that adaptation of S. frugiperda to SPI involves de novo synthesis and also up regulation of existing enzymes. Proteases from intestines of larvae reared on a diet with SPI showed insensitivity to the inhibitor. The proteases were also insensitive to a broad-spectrum potato proteinase inhibitor preparation. We propose that adaptation of S. frugiperda to SPI follows a "shotgun" approach, based on a general up regulation of a large set of endoproteinases.

While it has been well characterized that chemosensory receptors in guts of mammals have great influence on food preference, much remains elusive in insects. Insect chemosensory proteins (CSPs) are soluble proteins that could deliver chemicals to olfactory and gustatory receptors. Recent studies have identified a number of CSPs expressed in midgut in Lepidoptera insects, which started to reveal their roles in chemical recognition and stimulating appetite in midgut. In this study, we examined expression patterns in midgut of 21 Spodoptera litura CSPs (SlitCSPs) characterized from a previously reported transcriptome, and three CSPs were identified to be expressed highly in midgut. The orthologous relationships between midgut expressed CSPs in S. litura and those in Bombyx mori and Plutella xylostella also suggest a conserved pattern of CSP expression in midgut. We further demonstrated that the expression of midgut-CSPs may change in response to different host plants, and SlitCSPs could bind typical chemicals from host plant in vitro. Overall, our results suggested midgut expressed SlitCSPs may have functional roles, likely contributing to specialization and adaption to different ecosystems. Better knowledge of this critical component of the chemsensation signaling pathways in midguts may improve our understanding of food preference processes in a new perspective.

Plant volatiles serve as sensory cues for insects to find food and habitats. They are also reported to affect many insect behaviors. In the current study, we determined how exposures to taro (Colocasia esculenta) for 24h affect reproductive behaviors in Spodoptera litura. Further, we investigated the mechanisms that regulate taro volatiles-induced reproductive behavior in S. litura by recording peripheral, behavioral, and neuronal responses of male moths to sex pheromone components (Z9,E11-14:OAc and Z9,E12-14:OAc) and their mixtures in different ratios, as well as gene expression levels of sex pheromone receptors. The results showed that the exposure to taro volatiles significantly enhanced male mating rate, mating duration, and egg hatching rates, but not mating times and the number of oviposition per female. Consistently, the peripheral and behavioral responses of pre-exposed males to sex pheromone components and their mixtures wherein Z9,E11-14:OAc owned higher proportions, as well as neuronal responses to those at low dosages were significantly increased compared to non-exposed males. The expression levels of sex pheromone receptor genes were also significantly increased in pre-exposed males compared to non-exposed ones. These results suggest that taro volatile pre-exposures could promote the reproductive performance of S. litura by enhancing the competitiveness of male mating and communications between male and females. These findings provide new insights for the management of this insect pest as well as other moths.

The chitin synthase (CHS) gene B (4781 bp) of Spodoptera exigua (SeCHSB) was cloned by reverse-transcription PCR (RT-PCR) and 3'/5' RACE from the midgut. SeCHSB contains an open reading frame of 4572 nucleotides, encoding a protein of 1523 amino acids with a predicted molecular mass of approximately 174.6 kDa. Alignment of SeCHSB with class B CHSs of other insects showed a high degree of conservation in the putative catalytic domain region. The structure of the SeCHSB gene was analyzed and was found to be the same as that of Manduca sexta CHSB (MsCHSB), including 23 exons and 22 introns but without alternative exons. Southern blot analysis revealed that SeCHSB was a single copy gene and the presence of only two chitin synthase genes in S. exigua. Further investigation indicated that SeCHSB was specifically expressed in the midgut, and its transcript existed constitutively in the midgut from the 3rd instar larval stage to prepupae and reached highest expression on the 1st day of the fifth instar larval stage. These data suggest that SeCHSB is very important in midgut formation and development. Chitin synthase gene comparisons between different classes of insects using software tools revealed some interesting aspects of the similarity and divergence of the gene in the Class Insecta.

The present study revealed differentially expressed genes responsive to azadirachtin A (Aza) in Spodoptera litura cell line through suppression subtractive hybridization. In the Aza-responsive SSH library, approximately 270 sequences represent 53 different identified genes encoding proteins with various predicted functions, and the percentages of the gene clusters were 26.09% (genetic information processing), 11.41% (cell growth and death), 7.07% (metabolism), 6.52% (signal transduction/transport) and 2.72% (immunity), respectively. Eleven clones homologous to identified genes were selected to be confirmed through quantitative real time polymerase chain reaction. Among the eleven clones validated, all but one transcript of lipase showed an increase in SL cell line collected from ETA, whereas the transcripts of other genes were lower in the SL cell line collected from ETA compared with that of UETA. These genes were considered to be related to the response of SL cell line to Aza. These will provide a new clue to uncover the molecular mechanisms of Aza acting on SL cell line.

The induction of apoptosis by azadirachtin, a well-known botanical tetranortriterpenoid isolated from the neem tree (Azadirachta indica A. Juss) and other members of the Meliaceae, was investigated in Spodoptera frugiperda cultured cell line (Sf9). Morphological changes in Sf9 cells treated by various concentrations of azadirachtin were observed at different times under light microscopy. Morphological and biochemical analysis indicated that Sf9 cells treated by 1.5 μg/mL azadirachtin showed typical morphological changes, which were indicative of apoptosis and a clear DNA ladder. The flow cytometry analysis showed the apoptosis rate reached a maximum value of 32.66% at 24 h with 1.5 μg/mL azadirachtin in Sf9 cells. The inhibition of Sf9 cell proliferation suggested that the effect of azadirachtin was dose dependent and the EC50 at 48 and 72 h was 2.727 × 10(-6) and 6.348 × 10(-9) μg/mL, respectively. The treatment of azadirachtin in Sf9 cells could significantly increase the activity of Sf caspase-1, but showed no effect on the activity of Topo I, suggesting that the apoptosis induced by azadirachtinin Sf9 cells is through caspase-dependent pathway. These results provided not only a series of morphological, biochemical, and toxicological comprehensive evidences for induction of apoptosis by azadirachtin, but also a reference model for screening insect cell apoptosis inducers from natural compounds.

We used transient expression assays to assess the function of the baculovirus Spodoptera frugiperda M nucleopolyhedrovirus (SfMNPV) homologs of Autographa californica MNPV (AcMNPV) factors involved in late gene expression (lefs), in the Sf9 insect cell-line, which is permissive for both viruses. It is well-established that nineteen AcMNPV lefs support optimal levels of activity from a late promoter-reporter gene cassette in this assay. A subgroup of SfMNPV lefs predicted to function in transcription-specific events substituted the corresponding AcMNPV lefs very efficiently. When all SfMNPV lefs were assayed, including replication lefs, activity was low, but addition of two AcMNPV lefs not encoded in SfMNPV genome, resulted in augmented reporter activity. SfMNPV IE-1 was able to activate an early promoter cis-linked to an hr-derived element from SfMNPV but not from AcMNPV. However, the level of early promoter activation with SfMNPV IE-1 was lower compared to AcMNPV IE-1.

The tropical armyworm, Spodoptera litura (F.) (Lepidoptera: Noctuidae), is an important pest of tobacco, Nicotiana tabacum L. (Solanales: Solanaceae), in South China that is becoming increasingly resistant to pesticides. Six potential trap crops were evaluated to control S. litura on tobacco. Castor bean, Ricinus communis L. (Malpighiales: Euphorbiaceae), and taro, Colocasia esculenta (L.) Schott (Alismatales: Araceae), hosted significantly more S. litura than peanut, Arachis hypogaea L. (Fabales: Fabaceae), sweet potato, Ipomoea batata Lam. (Solanales: Convolvulaceae) or tobacoo in a greenhouse trial, and tobacco field plots with taro rows hosted significantly fewer S. litura than those with rows of other trap crops or without trap crops, provided the taro was in a fast-growing stage. When these crops were grown along with eggplant, Solanum melongena L. (Solanales: Solanaceae), and soybean, Glycines max L. (Fabales: Fabaceae), in separate plots in a randomized matrix, tobacco plots hosted more S. litura than the other crop plots early in the season, but late in the season, taro plots hosted significantly more S. litura than tobacco, soybean, sweet potato, peanut or eggplant plots. In addition, higher rates of S. litura parasitism by Microplitis prodeniae Rao and Chandry (Hymenoptera: Bracondidae) and Campoletis chlorideae Uchida (Ichnumonidae) were observed in taro plots compared to other crop plots. Although taro was an effective trap crop for managing S. litura on tobacco, it did not attract S. litura in the seedling stage, indicating that taro should either be planted 20-30 days before tobacco, or alternative control methods should be employed during the seedling stage.

Objective To evaluate the efficacy of Atalantia monophylla (A. monophylla) leaf in different solvent crude extracts and fractions against eggs of Spodoptera litura (S. litura). Methods Hexane, ethyl acetate and chloroform solvent extracts of A. monophylla leaf and 12 fractions from hexane extract were screened at 5.0%, 2.5%, 1.0% and 0.5% for crude extracts and 1 000, 500, 250 and 125 mg/kg for fractions against the eggs of S. litura for the ovicidal activity. LC50 and LC90 were calculated using probit analysis. Results Hexane crude extract showed maximum ovicidal activity of 61.94% at 5.0% concentration with a correlation value of r2=0.81, and least LC50 value of 3.06%. Hexane extract was fractionated using silica gel column chromatography and 12 fractions were obtained. Fraction 9 was active which showed maximum ovicidal activity of 75.61% at 1 000 mg/kg with the LC50 value of 318.65 mg/kg and LC90 value of 1 473.31 mg/kg. In linear regression analysis, significant and high correlation (r2=0.81%) was seen between concentration and ovicidal activity of hexane crude extracts and its active fraction. Conclusions As per our knowledge, this is the first report for ovicidal activity of A. monophylla against S. litura, A. monophylla could be used for the management of S. litura and other insect pests. PMID:23593580

Spodoptera frugiperda (Lepidoptera: Noctuidae) is a major pest in maize crops in Colombia, and affects several regions in America. A granulovirus isolated from S. frugiperda (SfGV VG008) has potential as an enhancer of insecticidal activity of previously described nucleopolyhedrovirus from the same insect species (SfMNPV). The SfGV VG008 genome was sequenced and analyzed showing circular double stranded DNA of 140,913 bp encoding 146 putative ORFs that include 37 Baculoviridae core genes, 88 shared with betabaculoviruses, two shared only with betabaculoviruses from Noctuide insects, two shared with alphabaculoviruses, three copies of own genes (paralogs) and the other 14 corresponding to unique genes without representation in the other baculovirus species. Particularly, the genome encodes for important virulence factors such as 4 chitinases and 2 enhancins. The sequence analysis revealed the existence of eight homologous regions (hrs) and also suggests processes of gene acquisition by horizontal transfer including the SfGV VG008 ORFs 046/047 (paralogs), 059, 089 and 099. The bioinformatics evidence indicates that the genome donors of mentioned genes could be alpha- and/or betabaculovirus species. The previous reported ability of SfGV VG008 to naturally co-infect the same host with other virus show a possible mechanism to capture genes and thus improve its fitness. PMID:25609309

The Vip3Aa protein is an insecticidal protein secreted by Bacillus thuringiensis during the vegetative stage of growth. The activity of this protein has been tested after different steps/protocols of purification using Spodoptera frugiperda as a control insect. The results showed that the Vip3Aa protoxin was stable and retained full toxicity after being subjected to common biochemical steps used in protein purification. Bioassays with the protoxin in S. frugiperda and S. exigua showed pronounced differences in LC(50) values when mortality was measured at 7 vs. 10d. At 7d most live larvae were arrested in their development. LC(50) values of "functional mortality" (dead larvae plus larvae remaining in the first instar), measured at 7d, were similar or even lower than the LC(50) values of mortality at 10d. This strong growth inhibition was not observed when testing the trypsin-activated protein (62 kDa) in either species. S. exigua was less susceptible than S. frugiperda to the protoxin form, with LC(50) values around 10-fold higher. However, both species were equally susceptible to the trypsin-activated form. Processing of Vip3Aa protoxin to the activated form was faster with S. frugiperda midgut juice than with S. exigua midgut juice. The results strongly suggest that the differences in the rate of activation of the Vip3Aa protoxin between both species are the basis for the differences in susceptibility towards the protoxin form.

Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae), the fall armyworm is the most economically important maize pest in the western hemisphere. This research focused on the genetic variability of the maize host strain because there is a lack of information in this area of S. frugiperda research. Amplified fragment length polymorphism (AFLP) was used to assess the genetic variability of S. frugiperda over a large geographic area. Twenty populations were collected from the maize, one population was collected from princess tree, one population was collected from lemon tree, and one population was collected from bermudagrass. The 23 populations were from Mexico, the continental United States, Puerto Rico, Brazil, and Argentina. The objective of this research was to evaluate whether the majority of genetic variability was within populations or between populations. The AFLP results showed that the majority of the genetic variability is within populations and not between populations, indicating minor gene flow and suggesting that S. frugiperda in the Western Hemisphere are an interbreeding population. PMID:20334595

The insect antifeedant anthraquinone aldehyde nordamnacanthal (1,3-dihydroxy-anthraquinone-2-al) was identified in Galium aparine L., and isolated from the root powder of akane (Rubia akane), a member of the Rubiaceae. Structure-activity relationship (SAR) studies using a series of anthraquinone analogues suggested that the aldehyde group on the anthraquinone was more important than the quinone moiety for antifeedant activity against the common cutworm (Spodoptera litura). High levels of nordamnacanthal were found in the seed leaf stage and in callus tissue induced from seedlings of G. aparine, but its concentration decreased with plant development. Since these compounds are natural pigments for dying textiles, we also evaluated the antifeedant activity against the carpet beetle (Attagenus japonicus ), a textile pest was also evaluated. While nordamnacanthal had strong antifeedant activity against the common cutworm, it did not show any antifeedant activity against the carpet beetle. The most effective antifeedant against the carpet beetle was the major constituent in the extract of R. trictorum, lucidin-3-O-primeveroside, a food pigment.

Median lethal concentrations (LC₅₀) were determined for four nematode populations (two strains of Steinernema feltiae, a S. feltiae hybrid, and S. bibionis) against fifth-instar fall armyworm (Spodoptera frugiperda) larvae and for the most virulent of these nematodes against different instars and stages of the insect. Based on lack of overlap of 95% fiducial limits, there were significant differences in virulence among the four nematodes. The LC₅₀ ranged from 7.6 to 33.3 nematodes/ 0.7 ml water, and slopes of the log dose-probit regression lines were similar except for the S. feltiae All strain. First-instar fall armyworms suffered virtually 100% mortality from the S. feltiae Mexican strain at 1.0 nematode/0.7 ml, and LC₅₀ were 2.3 and 7.9 nematodes/0.7 ml in third-instar and fifth-instar larvae, respectively. Pupae had 7-20% mortality at doses ranging from 30 to 60 nematodes/0.7 ml. PMID:19290189

The bacterium Bacillus thuringiensis produces Crystal (Cry) proteins that are toxic to a diverse range of insects. Transgenic crops that produce Bt Cry proteins are grown worldwide because of their improved resistance to insect pests. Although Bt “pyramid” cotton that produces both Cry1A and Cry2A is predicted to be more resistant to several lepidopteran pests, including Spodoptera exigua, than plants that produce Cry1Ac alone, the mechanisms responsible for the toxicity of Cry2Aa in S. exigua are not well understood. We identified several proteins that bind Cry2Aa (polycalin, V-ATPase subunits A and B, actin, 4-hydroxybutyrate CoA-transferase [4-HB-CoAT]), and a receptor for activated protein kinase C (Rack), in S. exigua. Recombinant, expressed versions of these proteins were able to bind the Cry2Aa toxin in vitro assays. RNA interference gene knockdown of the Se-V-ATPase subunit B significantly decreased the susceptibility of S. exigua larvae to Cry2Aa, whereas knockdown of the other putative binding proteins did not. Moreover, an in vitro homologous competition assay demonstrated that the Se-V-ATPase subunit B binds specifically to the Cry2Aa toxin, suggesting that this protein acts as a functional receptor of Cry2Aa in S. exigua. This the first Cry2Aa toxin receptor identified in S. exigua brush-border membrane vesicles. PMID:28067269

Although the general pathway of sex pheromone synthesis in moth species has been established, the molecular mechanisms remain poorly understood. The common cutworm Spodoptera litura is an important agricultural pest worldwide and causes huge economic losses annually. The female sex pheromone of S. litura comprises Z9,E11-14:OAc, Z9,E12-14:OAc, Z9-14:OAc, and E11-14:OAc. By sequencing and analyzing the transcriptomic data of the sex pheromone glands, we identified 94 candidate genes related to pheromone biosynthesis (55 genes) or chemoreception (39 genes). Gene expression patterns and phylogenetic analysis revealed that two desaturase genes (SlitDes5 and SlitDes11) and one fatty acyl reductase gene (SlitFAR3) showed pheromone gland (PG) biased or specific expression, and clustered with genes known to be involved in pheromone synthesis in other moth species. Furthermore, 4 chemoreception related genes (SlitOBP6, SlitOBP11, SlitCSP3, and SlitCSP14) also showed higher expression in the PG, and could be additional candidate genes involved in sex pheromone transport. This study provides the first solid background information that should facilitate further elucidation of sex pheromone biosynthesis and transport, and indicates potential targets to disrupt sexual communication in S. litura for a novel pest management strategy.

Presents a digest of basic developmental information about children's feeding skills and behaviors, and gives general feeding recommendations. Also addresses requirements for feeding children with developmental disabilities and chronic medical conditions for which adapted environments or monitored nutrient intake may be necessary. (ET)

The chemical composition of the soybean pods armyworm Spodoptera cosmioides (Walker, 1858) (Lepidoptera: Noctuidae) and Anticarsia gemmatalis Hübner, 1818 (Lepidoptera: Erebidae) larval cuticles was evaluated using gas chromatography coupled to a mass detector (GC-MS). Among the usual lipids found in the insect cuticle, α-tocopherol and α-tocopheryl acetate were also isolated from S. cosmioides. On the other hand, no vitamin E derivative was found in A. gemmatalis exuvia. This is the first report of vitamin E occurrence in the insect's cuticle.

Beet armyworm, Spodoptera exigua (Hübner), is a serious pest of vegetables in China, and its control is heavily dependent on chemical insecticides. The current resistance status of nine insecticides was investigated in 16 field populations collected from seven provinces of China during 2009-2012. Compared with the susceptible strain WH-S, some field populations evolved various levels of resistance to eight of the nine insecticides tested: emamectin benzoate (4- to 348-fold), indoxacarb (2- to 41-fold), spinosad (5- to 38-fold), chlorantraniliprole (2- to 44-fold), tebufenozide (2- to 87-fold), chlorfluazuron (3- to 31-fold), cypermethrin (79- to 1240-fold), and chlorpyrifos (8- to 3,080-fold), but no significant resistance was detected to chlorfenapyr (0.4- to 7-fold). This indicates that chlorfenapyr has no cross-resistance with these other currently used insecticides. Four consecutive years' resistance screening at two places shows that resistance patterns were different between populations from Luhe (Jiangsu Province) and Fengxian (Shanghai), which are approximately 300 km apart. Resistance levels to chlorpyrifos were much higher in populations from Luhe (877- to 3,080-fold) than from Fengxian (8- to 110-fold). Fengxian populations developed moderate levels of resistance to tebufenozide (13- to 87-fold), but no resistance in Luhe populations (2- to 6-fold). However, Luhe populations developed moderate levels of resistance to chlorfluazuron (21- to 31-fold), but there was no resistance in Fengxian populations (3- to 5-fold). It is suggested that local insecticide selection determined resistance patterns although S. exigua has long-distance migratory potential. Adaptive resistance management tactics (such as rotations) should be designed and implemented based on the resistance patterns of S. exigua for each geographic area.

Spodoptera litura (F.) causes considerable economic damage to multiple agro-crops annually in many countries. In this study, the demography of S. litura reared on cabbage and taro was investigated using the age-stage, two-sex life table at 25±1°C, 60±10% relative humidity, and a photoperiod of 12: 12 (L:D) h. Our results showed that the net reproductive rate, intrinsic rate, and finite rate of population increase on cabbage (1893.1 offspring, 0.2374 d(−1), and 1.2679 d(−1)) were all not significantly different from those on taro (1361.0 offspring, 0.2298 d(−1), and 1.2584 d(−1)). The net consumption rate on cabbage (439.1 cm2) was, however, three times higher than that on taro (141.7 cm2). According to the population parameters, both cabbage and taro are suitable host plants for S. litura. When both the population growth rate and the consumption rate were taken into consideration, the finite consumption rate on cabbage (ω=3.8054) was significantly higher than that on taro (ω=1.3184). In Taiwan, taro and cabbage are commonly planted in adjacent farm plots, with taro being grown from March to November and cabbage from October to April. Because of the overlapping growth periods of the two crops, S. litura can easily propagate throughout the year by switching between the adjacent crops during the overlap periods. Pest management strategies for controlling S. litura must be thoroughly reevaluated based on ecological characteristics, including its life table and consumption rate on its major host plants.

Metal hyperaccumulation may be an elemental defense, in which high concentrations of a metal in plant tissues decrease herbivore survival or growth rate. The Joint Effects Hypothesis suggests that a combination of metals, or a combination of a metal with an organic compound, may have an enhanced defensive effect. The enhancement may be additive or synergistic: in either case the concentration of a particular metal necessary to provide a defensive benefit for the plant is lowered. We tested the Joint Effects Hypothesis using Spodoptera exigua (beet armyworm) neonates fed artificial diets. Metal + metal experiments utilized diets amended with metal pairs, using four metals commonly hyperaccumulated by plants (Co, Cu, Ni, and Zn). We also conducted metal + organic compound experiments, pairing each metal with nicotine, mustard seed powder, or tannic acid. We tested for joint effects using both lethal (LC20 levels) and sublethal concentrations (10-25 % reduced larval weight) of the chemicals tested. For all experiments, either additive or synergistic effects were found. Of the metal + metal pairs tested, three (Co + Cu, Cu + Zn, and Ni + Zn) were synergistic in lethal concentration tests and only Co + Cu was synergistic in sublethal tests. For metal + organic combination lethal tests, synergism occurred for all combinations except for Co or Ni + nicotine, Ni + mustard seed powder, and Zn + nicotine. For sublethal tests, Zn + all three organic chemicals, Co + mustard seed powder or tannic acid, and Cu + nicotine, were synergistic. These results support the Joint Effects Hypothesis, suggesting that metals combined with other metals or organic compounds may be more effective against herbivores than individual metals.

To develop a better understanding of the natural distribution of the fall armyworm, Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae), and to update the knowledge of the incidence of its complex of parasitoids. S. frugiperda, samplings in whorl-stage corn were carried out in provinces of Argentina from 1999 to 2003. S. frugiperda larvae were collected from corn in localities of the provinces of Tucumán, Salta, Jujuy, Santiago del Estero, La Rioja, Córdoba, San Luis, Chaco and Misiones. In each locality 30 corn plants were sampled and only larvae located in those plants were collected. The parasitoids that emerged from S. frugiperda larvae were identified and counted. The abundance of the parasitoids and the parasitism rate were estimated. The S. frugiperda parasitoids collected were Campoletis grioti (Blanchard) (Hymenoptera: Ichneumonidae), Chelonus insularis (Cresson) (Hymenoptera: Braconidae), Archytas marmoratus (Townsend) (Diptera Tachinidae) and/or A. incertus (Macquart), Ophion sp. (Hymenoptera: Ichneumonidae), Euplectrus platyhypenae Howard (Hymenoptera: Eulophidae), and Incamyia chilensis (Aldrich) (Diptera Tachinidae). C. grioti was the most abundant and frequent during the five-year survey. Similar diversity of parasitoids was obtained in all the provinces, with the exception of I. chilensis and E. platyhypenae that were recovered only in the province of Salta. In the Northwestern region, in Tucumán, C. grioti and species of Archytas were the most abundant and frequent parasitoids. On the contrary, in Salta and Jujuy Ch. insularis was the parasitoid most abundant and frequently recovered. The parasitism rate obtained in Tucumán, Salta and Jujuy provinces were 21.96%, 17.87% and 6.63% respectively with an average of 18.93%. These results demonstrate that hymenopteran and dipteran parasitoids of S. frugiperda occurred differentially throughout the Argentinian provinces and played an important role on the natural control of the S. frugiperda larval

Glycosylasparaginase (EC 3.5.1.26) from Sf9 cells (Spodoptera frugiperda) was purified to homogeneity with a specific activity of 2.1 unit/mg. The enzyme is composed of two non-identical alpha/beta subunits joined by strong non-covalent forces and has one glycosylation site located in the alpha subunit. Molecular masses of the subunits were determined to be 28 kDa and 17 kDa by SDS-PAGE. Native enzyme existed in quaternary structures of either heterodimer (alpha beta) or heterotetramer (alpha 2 beta 2). These forms exhibited different ionic characteristics during DE52 anion exchange chromatography, and their molecular masses were determined to be 47 kDa and 101 kDa by gel filtration. The enzyme was thermostable, requiring 65-70 degrees C to be denatured, and it had a broad pH optimum from 4-10.5 with a pKa around 6. SDS easily inactivated the enzyme. The K(m) of glycosylasparaginase for its normal substrate GlcNAc-Asn was 0.88 mM. The enzyme also exhibited asparaginase activity with a K(m) of 3.0 mM for asparagine. N-terminal amino acids of the denatured subunits were sequenced and degenerate primers were designed for cloning its cDNA using PCR and 5' and 3' RACE. Glycosylasparaginase cDNAs from bovine and rat were also cloned using similar strategies, and primary structures of glycosylasparaginases from six species (human, bovine, rat, mouse, Sf9 cells and Flavobacterium) have been compared and related to a recent crystal structure of the human enzyme.

The common cutworm, Spodoptera litura (F.), is a serious crop pest with a strong migratory ability. Previous studies on the migration of S. litura were mostly carried out in its "year-round breeding region" (YBR) or "overwintering region" (OR). However, the pattern of seasonal movements in its "summer breeding region" (SBR; i.e., northern China where they cannot overwinter) remains unknown. Here, we present data from an 11-yr study of this species made by searchlight trapping on Beihuang (BH) Island in the center of the Bohai Strait, which provides direct evidence that S. litura regularly migrates across this sea. There was considerable yearly and monthly variation in the number of S. litura trapped on BH, with the vast majority trapped in the autumn. The mean time from the earliest trapping to the latest trapping within a year was 110±12 d during 2003-2013, with the shortest time span of 40 d in 2003 and the longest of 166 d in 2012. S. litura moths had downwind displacement rather than randomly by heading toward their seasonally favorable direction (i.e. toward southwest in the four autumn migration events by prevailing northeasterly winds). Some females trapped in July showed a relatively higher proportion of having mated and a degree of ovarian development, suggesting that the migration of this species is not completely bound by the "oogenesis-flight syndrome." These findings provide a good starting point of research on S. litura migration between its OR (or YBR) and SBR, which will help us develop more effective regional management strategies against this pest.

The common cutworm, Spodoptera litura, is a rapidly reproducing pest of numerous agricultural ecosystems worldwide. The use of pesticides remains the primary means for controlling S. litura, despite their negative ecological impact and potential threat to human health. The use of exogenous hormone analogs may represent an alternative to insecticides. Juvenile hormones (JHs) play an important role in the reproductive systems of female insects, but the effects of pyriproxyfen, a JH analog, on reproduction in S. litura were poorly understood. In this paper, we topically treated the newly emerged females with 20, 60, or 100 μg of pyriproxyfen to determine its effects on reproduction. Then, we examined the expression of vitellogenin (Vg) and three hormone receptors, USP, HR3, and EcR, using quantitative reverse transcription and real-time polymerase chain reaction (qRT-PCR), and found that pyriproxyfen up-regulated the expression of Vg, USP, and HR3, whereas the expression of EcR was unaffected. An analysis of fecundity showed that the peak oviposition day, lifespan, and oviposition period were progressively shortened as the pyriproxyfen dosage increased. We also found that pyriproxyfen decreased egg laying amount, whereas the number of mature eggs that remained in the ovarioles of dead females increased as the pyriproxyfen dosage increased. We examined oocytes using transmission electron microscopy and found that treatment with 100 μg of pyriproxyfen increased the metabolism by increasing the amount of rough endoplasmic reticulum and mitochondria in the primary oocytes. Our results suggest that the topical application of pyriproxyfen on newly emerged females can efficiently reduce reproduction in S. litura and may represent an alternative to the use of insecticides for controlling the agricultural pest.

The effects of extracts of sesame, Sesamum indicum L. (Liamiales: Pedaliaceae), and whole leaves of some selected cultivars of sesame were tested using a natural host Spodoptera litura (F.) (Lepidoptera: Noctuidae). Indices taken using the immature stages include; diet utilization, growth and development and induction of detoxification enzymes. The results indicate that S. litura generally selects its food amongst cultivars within 6 hours after food presentation. Growth and development of the insect is controlled also by plant acceptability and quality. Although all the cultivars tested significantly limit insect growth and development the variety 56S-radiatum did not allow a complete life cycle as pupation from first instar stage was 0%. Generally the crucial period for immature S. litura was the larval period, especially the first two instars where the weight of an insect fed on an experimental diet was three times lower than that of a control diet. The larval developmental period was greater than 40 days as compared to 17 days for insects fed a control diet. S. litura also had lowered efficiency in utilizing ingested food, from a low of 13% in a sesame cultivar to 45% in the control diet. The key detoxification enzyme was a glutathione s-transferase that was confirmed by a 6-fold increase between S. litura fed a plant cultivar vs. a control diet towards the substrate 1,2-dichloro-4-nitrobenzene. First and second instars of S. litura have a relatively reduced detoxification of enzymes in response to plant cultivar diets leading to low survival. A 3% v/w crude extract of the cultivars increased enzyme induction towards all the tested substrates. PMID:20050772

MicroRNAs (miRNAs) as small non-coding RNAs play important roles in many biological processes such as development, cell signalling and immune response. Studies also suggest that miRNAs are important in host-virus interactions where the host limits virus infection by differentially expressing miRNAs that target essential viral genes. Here, we identified conserved and new miRNAs from Spodoptera frugiperda cells (Sf9) using a combination of deep sequencing and bioinformatics as well as experimental approaches. S. frugiperda miRNAs share common features of miRNAs in other organisms, such as uracil (U) at the 5' end of miRNA. The 5' ends of the miRNAs were more conserved than the 3' ends, revealing evolutionary protection of the seed region in miRNAs. The predominant miRNAs were found to be conserved among arthropods. The majority of homologous miRNAs were found in Bombyx mori, with 76 of the 90 identified miRNAs. We found that seed shifting and arm switching have happened in this insect's miRNAs. Expression levels of the majority of miRNAs changed following baculovirus infection. Results revealed that baculovirus infection mainly led to an overall suppression of cellular miRNAs. We found four different genes being regulated by sfr-miR-184 at the post-transcriptional level. The data presented here further support conservation of miRNAs in insects and other organisms. In addition, the results reveal a differential expression of host miRNAs upon baculovirus infection, suggesting their potential roles in host-virus interactions. Seed shifting and arm switching happened during evolution of miRNAs in different insects and caused miRNA diversification, which led to changes in the target repository of miRNAs.

The tropical armyworm, Spodoptera litura (F.) (Lepidoptera: Noctuidae), is an important pest of tobacco, Nicotiana tabacum L. (Solanales: Solanaceae), in South China that is becoming increasingly resistant to pesticides. Six potential trap crops were evaluated to control S. litura on tobacco. Castor bean, Ricinus communis L. (Malpighiales: Euphorbiaceae), and taro, Colocasia esculenta (L.) Schott (Alismatales: Araceae), hosted significantly more S. litura than peanut, Arachis hypogaea L. (Fabales: Fabaceae), sweet potato, Ipomoea batata Lam. (Solanales: Convolvulaceae) or tobacoo in a greenhouse trial, and tobacco field plots with taro rows hosted significantly fewer S. litura than those with rows of other trap crops or without trap crops, provided the taro was in a fast-growing stage. When these crops were grown along with eggplant, Solanum melongena L. (Solanales: Solanaceae), and soybean, Glycines max L. (Fabales: Fabaceae), in separate plots in a randomized matrix, tobacco plots hosted more S. litura than the other crop plots early in the season, but late in the season, taro plots hosted significantly more S. litura than tobacco, soybean, sweet potato, peanut or eggplant plots. In addition, higher rates of S. litura parasitism by Microplitis prodeniae Rao and Chandry (Hymenoptera: Bracondidae) and Campoletis chlorideae Uchida (Ichnumonidae) were observed in taro plots compared to other crop plots. Although taro was an effective trap crop for managing S. litura on tobacco, it did not attract S. litura in the seedling stage, indicating that taro should either be planted 20–30 days before tobacco, or alternative control methods should be employed during the seedling stage. PMID:20874598

A Nicaraguan field isolate (SfNIC) of Spodoptera frugiperda nucleopolyhedrovirus was purified by plaque assay on Sf9 cells. Nine distinct genotypes, A to I, were identified by their restriction endonuclease profiles. Variant SfNIC-B was selected as the standard because its restriction profile corresponded to that of the wild-type isolate. Physical maps were generated for each of the variants. The differences between variants and the SfNIC-B standard were confined to the region between map units 9 and 32.5. This region included PstI-G, PstI-F, PstI-L, PstI-K and EcoRI-L fragments. Eight genotypes presented a deletion in their genome compared with SfNIC-B. Occlusion body-derived virions of SfNIC-C, -D and -G accounted for 41% of plaque-purified clones. These variants were not infectious per os but retained infectivity by injection into S. frugiperda larvae. Median 50% lethal concentration values for the other cloned genotypes were significantly higher than that of the wild type. The variants also differed in their speed of kill. Noninfectious variants SfNIC-C and -D lacked the pif and pif-2 genes. Infectivity was restored to these variants by plasmid rescue with a plasmid comprising both pif and pif-2. Transcription of an SfNIC-G gene was detected by reverse transcription-PCR in insects, but no fatal disease developed. Transcription was not detected in SfNIC-C or -D-inoculated larvae. We conclude that the SfNIC population presents high levels of genetic diversity, localized to a 17-kb region containing pif and pif-2, and that interactions among complete and deleted genotypic variants will likely influence the capacity of this virus to control insect pests. PMID:15345446

The DNA restriction map for the enzymes BamHI, BglII, PstI, and XbaI of SeMNPV-US (Se-US), the best-studied Nucleopolyhedrovirus strain isolated from Spodoptera exigua in California, was used as a reference to construct that of SeMNPV-SP2 (Se-SP2), a closely related Spanish strain of the same virus. After coinfection of S. exigua with both the Se-US and Se-SP2 strains per os, a recombinant virus (SeMNPV-SUR1 [Se-SUR1]) was detected after one passage which quickly replaced the parental viruses. A physical map of Se-SUR1 DNA was constructed for BamHI, BglII, PstI, and XbaI and compared to that of the parental viruses, Se-US and Se-SP2. Se-SUR1 is the result of at least four crossover events between Se-US and Se-SP2 and not of selection of a minor variant in one of the parental viruses. Bioassays of the recombinant and parental strains against L2 beet armyworm larvae demonstrated that their 50% lethal dose values were not significantly different. The differences in median lethal time values are too small to explain the replacement of the parental viruses by the recombinant virus upon successive passage in vivo, although it cannot be ruled out as an explanation for the selective advantage of the recombinant strain, Se-SUR1. The consequences of the release of nonindigenous or recombinant baculovirus strains in agro-ecosystems are discussed. PMID:9251191

The common cutworm, Spodoptera litura, is a rapidly reproducing pest of numerous agricultural ecosystems worldwide. The use of pesticides remains the primary means for controlling S. litura, despite their negative ecological impact and potential threat to human health. The use of exogenous hormone analogs may represent an alternative to insecticides. Juvenile hormones (JHs) play an important role in the reproductive systems of female insects, but the effects of pyriproxyfen, a JH analog, on reproduction in S. litura were poorly understood. In this paper, we topically treated the newly emerged females with 20, 60, or 100 μg of pyriproxyfen to determine its effects on reproduction. Then, we examined the expression of vitellogenin (Vg) and three hormone receptors, USP, HR3, and EcR, using quantitative reverse transcription and real-time polymerase chain reaction (qRT-PCR), and found that pyriproxyfen up-regulated the expression of Vg, USP, and HR3, whereas the expression of EcR was unaffected. An analysis of fecundity showed that the peak oviposition day, lifespan, and oviposition period were progressively shortened as the pyriproxyfen dosage increased. We also found that pyriproxyfen decreased egg laying amount, whereas the number of mature eggs that remained in the ovarioles of dead females increased as the pyriproxyfen dosage increased. We examined oocytes using transmission electron microscopy and found that treatment with 100 μg of pyriproxyfen increased the metabolism by increasing the amount of rough endoplasmic reticulum and mitochondria in the primary oocytes. Our results suggest that the topical application of pyriproxyfen on newly emerged females can efficiently reduce reproduction in S. litura and may represent an alternative to the use of insecticides for controlling the agricultural pest. PMID:26444432

Evolution of insect resistance to transgenic crops containing Bacillus thuringiensis (Bt) genes is a serious threat to the sustainability of this technology. However, field resistance related to the reduced efficacy of Bt maize has not been documented in any lepidopteran pest in the mainland U.S. after 18 years of intensive Bt maize planting. Here we report compelling evidence of field resistance in the fall armyworm, Spodoptera frugiperda (J.E. Smith), to Cry1F maize (TC 3507) in the southeastern region of the U.S. An F2 screen showed a surprisingly high (0.293) Cry1F resistance allele frequency in a population collected in 2011 from non-Bt maize in south Florida. Field populations from non-Bt maize in 2012–2013 exhibited 18.8-fold to >85.4-fold resistance to purified Cry1F protein and those collected from unexpectedly damaged Bt maize plants at several locations in Florida and North Carolina had >85.4-fold resistance. In addition, reduced efficacy and control failure of Cry1F maize against natural populations of S. frugiperda were documented in field trials using Cry1F-based and pyramided Bt maize products in south Florida. The Cry1F-resistant S. frugiperda also showed a low level of cross-resistance to Cry1A.105 and related maize products, but not to Cry2Ab2 or Vip3A. The occurrence of Cry1F resistance in the U.S. mainland populations of S. frugiperda likely represents migration of insects from Puerto Rico, indicating the great challenges faced in achieving effective resistance management for long-distance migratory pests like S. frugiperda. PMID:25401494

Evolution of insect resistance to transgenic crops containing Bacillus thuringiensis (Bt) genes is a serious threat to the sustainability of this technology. However, field resistance related to the reduced efficacy of Bt maize has not been documented in any lepidopteran pest in the mainland U.S. after 18 years of intensive Bt maize planting. Here we report compelling evidence of field resistance in the fall armyworm, Spodoptera frugiperda (J.E. Smith), to Cry1F maize (TC 3507) in the southeastern region of the U.S. An F2 screen showed a surprisingly high (0.293) Cry1F resistance allele frequency in a population collected in 2011 from non-Bt maize in south Florida. Field populations from non-Bt maize in 2012-2013 exhibited 18.8-fold to >85.4-fold resistance to purified Cry1F protein and those collected from unexpectedly damaged Bt maize plants at several locations in Florida and North Carolina had >85.4-fold resistance. In addition, reduced efficacy and control failure of Cry1F maize against natural populations of S. frugiperda were documented in field trials using Cry1F-based and pyramided Bt maize products in south Florida. The Cry1F-resistant S. frugiperda also showed a low level of cross-resistance to Cry1A.105 and related maize products, but not to Cry2Ab2 or Vip3A. The occurrence of Cry1F resistance in the U.S. mainland populations of S. frugiperda likely represents migration of insects from Puerto Rico, indicating the great challenges faced in achieving effective resistance management for long-distance migratory pests like S. frugiperda.

To develop a better understanding of the natural distribution of the fall armyworm, Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae), and to update the knowledge of the incidence of its complex of parasitoids. S. frugiperda, samplings in whorl-stage corn were carried out in provinces of Argentina from 1999 to 2003. S. frugiperda larvae were collected from corn in localities of the provinces of Tucumán, Salta, Jujuy, Santiago del Estero, La Rioja, Córdoba, San Luis, Chaco and Misiones. In each locality 30 corn plants were sampled and only larvae located in those plants were collected. The parasitoids that emerged from S. frugiperda larvae were identified and counted. The abundance of the parasitoids and the parasitism rate were estimated. The S. frugiperda parasitoids collected were Campoletis grioti (Blanchard) (Hymenoptera: Ichneumonidae), Chelonus insularis (Cresson) (Hymenoptera: Braconidae), Archytas marmoratus (Townsend) (Diptera Tachinidae) and/or A. incertus (Macquart), Ophion sp. (Hymenoptera: Ichneumonidae), Euplectrus platyhypenae Howard (Hymenoptera: Eulophidae), and Incamyia chilensis (Aldrich) (Diptera Tachinidae). C. grioti was the most abundant and frequent during the five-year survey. Similar diversity of parasitoids was obtained in all the provinces, with the exception of I. chilensis and E. platyhypenae that were recovered only in the province of Salta. In the Northwestern region, in Tucumán, C. grioti and species of Archytas were the most abundant and frequent parasitoids. On the contrary, in Salta and Jujuy Ch. insularis was the parasitoid most abundant and frequently recovered. The parasitism rate obtained in Tucumán, Salta and Jujuy provinces were 21.96%, 17.87% and 6.63% respectively with an average of 18.93%. These results demonstrate that hymenopteran and dipteran parasitoids of S. frugiperda occurred differentially throughout the Argentinian provinces and played an important role on the natural control of the S. frugiperda larval

The pheromone composition of the Spanish population of the beet armyworm (BAW), Spodoptera exigua (Lepidoptera: Noctuidae), was identified. Analysis of female gland extracts showed the presence of compounds Z9,E12-14:Ac (1), Z9-14:Ac (2), Z11-16:Ac (3), Z9,E12-14:OH (4), Z9-14:OH (5), and Z11-16:OH (6) in a ratio of 26:11:1:22:31:9. The amount of compound per gland ranged from 2.08 ng for 5 to 0.09 ng for 3. However, analysis of female volatiles by SPME revealed only the presence of compounds, 1, 2, 3, and 5 in a 34:40:4:22 ratio. In electroantennogram assays, compound 1 elicited the highest response, and the C14 acetates evoked higher electrophysiological responses than the corresponding alcohols or C16 isomers. In a wind tunnel, no behavioral difference was observed between formulations based on the gland extracts and female volatiles. In both cases, males responded as when virgin females were used as the attractant source. Compound 1 alone elicited upwind flight by males, but required the presence of compound 5 in a 80:20 to 40:60 ratio for full activity. Ternary mixtures of 1, 5 and the minor components did not improve the performance of the 1+5 blend in a 60:40 ratio. In the field, the mixture 1+5+3 in a 56:37:7 ratio was the most attractive formulation, and is expected to be useful in future pest control strategies.

Insect chemosensory genes have been considered as potential molecular targets to develop alternative strategies for pest control. However, in Spodoptera exigua, a seriously polyphagous agricultural pest, only a small part of such genes have been identified and characterized to date. Here, using a bioinformatics screen a total of 79 chemosensory genes were identified from a public transcriptomic data of different developmental stages (eggs, 1st to 5th instar larvae, pupae, female and male adults), including 34 odorant binding proteins (OBPs), 20 chemosensory proteins (CSPs), 22 chemosensory receptors (10 odorant receptors (ORs), six gustatory receptors (GRs) and six ionotropic receptors (IRs)) and three sensory neuron membrane proteins (SNMPs). Notably, a new group of lepidopteran SNMPs (SNMP3 group) was found for the first time in S. exigua, and confirmed in four other moth species. Further, reverse transcription PCR (RT-PCR) and quantitative real time PCR (qPCR) were employed respectively to validate the sequences and determine the expression patterns of 69 identified chemosensory genes regarding to sexes, tissues and stages. Results showed that 67 of these genes could be detected and reconstructed in at least one tissue tested. Further, 60 chemosensory genes were expressed in adult antennae and 52 in larval heads with the antennae, whereas over half of the genes were also detected in non-olfactory tissues like egg and thorax. Particularly, S. exigua OBP2 showed a predominantly larval head-biased expression, and functional studies further indicated its potentially olfactory roles in guiding food searching of larvae. This work suggests functional diversities of S. exigua chemosensory genes and could greatly facilitate the understanding of olfactory system in S. exigua and other lepidopteran species.

In the preparation of the gaseous reactant feed to undergo a chemical reaction requiring the presence of steam, the efficiency of overall power utilization is improved by premixing the gaseous reactant feed with water and then heating to evaporate the water in the presence of the gaseous reactant feed, the heating fluid utilized being at a temperature below the boiling point of water at the pressure in the volume where the evaporation occurs.

and feed forward stabilization) have been implemented. An on-mount gyro system consists of gyroscopes mounted on the radar antenna which sense...antenna motion and send compensating signals back to the antenna servo mechanism. Feed forward stabilization consists of determining antenna angular rates...caused by ships attitude changes, as measured by a stable platform (such as SINS), and feeding compensating signals back to the antenna servo

The fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), is a target of transgenic maize and cotton expressing Bacillus thuringiensis (Bt) proteins in both North and South America. In 2013 and 2014, a total of 215 F2 two-parent families of S. frugiperda were established usin...

Cytochrome P450 monooxygenases (P450) play a prominent role in the adaptation of insects to host plant chemical defenses. To investigate the potential role of P450s in adaptation of the lepidopteran pest Spodoptera litura to host plant allelochemicals, an expressed sequence data set derived from 6th...

This is the first study to report a high-throughput approach integrating gene expression data from Spodoptera frugiperda guts and their associated metatranscriptomes. Our datasets provide information on the potential effects of environmental conditions on the expression profile of S. frugiperda larval guts, their associated metatranscriptome, and putative interactions between them. PMID:26184938

VLBI2010 requires a feed that simultaneously has high efficiency over the full 2.2-14 GHz frequency range. The simultaneity requirement implies that the feed must operate at high efficiency over the full frequency range without the need to adjust its focal position to account for frequency dependent phase centre variations. Two feeds meet this specification: The Eleven Feed developed at Chalmers University. (For more information, contact Miroslav Pantaleev, miroslav.pantaleev@chalmers.se. The Eleven Feed, integrated with LNA's in a cryogenic receiver, is available as a product from Omnisys Instruments, info@omnisys.se). The Quadruple Ridged Flared Horn (QRFH) developed at the California Institute of Technology. (For more information please contact Ahmed Akgiray, aakgiray@ieee.org or Sander Weinreb, sweinreb@caltech.edu) Although not VLBI2010 compliant, two triband S/X/Ka feeds are also being developed for the commissioning of VLBI2010 antennas, for S/X observations during the VLBI2010 transition period, and to support X/Ka CRF observations. The two feeds are: The Twin Telescopes Wettzell (TTW) triband feed developed by Mirad Microwave. (For more information please contact Gerhard Kronschnabl, Gerhard.Kronschnabl@bkg.bund.de) The RAEGE (Spain) triband feed developed at Yebes Observatory. (For more information please contact Jose Antonio Lopez Perez, ja.lopezperez@oan.es)

An infectious waste feed system for comminuting infectious waste and feeding the comminuted waste to a combustor automatically without the need for human intervention. The system includes a receptacle for accepting waste materials. Preferably, the receptacle includes a first and second compartment and a means for sealing the first and second compartments from the atmosphere. A shredder is disposed to comminute waste materials accepted in the receptacle to a predetermined size. A trough is disposed to receive the comminuted waste materials from the shredder. A feeding means is disposed within the trough and is movable in a first and second direction for feeding the comminuted waste materials to a combustor.

Vegetative insecticidal protein (Vip3A) is synthesized as an extracellular insecticidal toxin by certain strains of Bacillus thuringiensis. Vip3A is active against several lepidopteran pests of crops. Polyphagous pest, Spodoptera frugiperda, and its cell line Sf21 are sensitive for lyses to Vip3A. Screening of cDNA library prepared from Sf21 cells through yeast two-hybrid system with Vip3A as bait identified ribosomal protein S2 as a toxicity-mediating interacting partner protein. The Vip3A-ribosomal-S2 protein interaction was validated by in vitro pulldown assays and by RNA interference-induced knockdown experiments. Knockdown of expression of S2 protein in Sf21 cells resulted in reduced toxicity of the Vip3A protein. These observations were further extended to adult fifth-instar larvae of Spodoptera litura. Knockdown of S2 expression by injecting corresponding double-stranded RNA resulted in reduced mortality of larvae to Vip3A toxin. Intracellular visualization of S2 protein and Vip3A through confocal microscopy revealed their interaction and localization in cytoplasm and surface of Sf21 cells.

"Feeding up" establishes a substantive line of inquiry that compels learners to engage in investigation and inquire. It also forms the basis for the assessments that follow. Once students understand the purpose and begin to work, they receive "feedback" that is timely and scaffolds their understanding. Based on their responses, the teacher gains a…

By-product feeds are generated from the production of food, fiber, and bio-energy products for human consumption. They include plant feedstuffs such as hulls, stalks, peels, and oil seed meals, and animal by-products such as blood meal, fats, bone meal, or processed organ meats. Some feed by-product...

This paper has two major purposes: first, to consider how infant feeding behavior may fit into attachment theory; and second, to cite some evidence to show how an infant's early interaction with his mother in the feeding situation is related to subsequent development. It was found that sucking and rooting are precursor attachment behaviors that…

Human salmonellosis outbreaks have been linked to contamination of animal feeds. Thus it is crucial to employ sensitive Salmonella detection methods for animal feeds. Based on a review of the literature, Salmonella sustains acid injury at about pH 4.0 to5.0. Low pH can also alter the metabolism of S...

The journey children make from tube feeding to oral feeding is personal for each child and family. There is a sequence of predictable plateaus that children climb as they move toward orally eating. By better understanding this sequence, parents and children can maximize the development, learning, enjoyment and confidence at each plateau. The…

Feeding is essential for survival, whereas withdrawal and escape reactions are fundamentally protective. These critical behaviors can compete for an animal's resources when an acutely painful stimulus affects the animal during feeding. One solution to the feeding-withdrawal conflict is to optimize feeding by suppressing pain. We examined whether rats continue to feed when challenged with a painful stimulus. During feeding, motor withdrawal responses to noxious paw heat either did not occur or were greatly delayed. To investigate the neural basis of sensory suppression accompanying feeding, we recorded from brainstem pain-modulatory neurons involved in the descending control of pain transmission. During feeding, pain-facilitatory ON cells were inhibited and pain-inhibitory OFF cells were excited. When a nonpainful somatosensory stimulus preactivated ON cells and preinhibited OFF cells, rats interrupted eating to react to painful stimuli. Inactivation of the brainstem region containing ON and OFF cells also blocked pain suppression during eating, demonstrating that brainstem pain-modulatory neurons suppress motor reactions to external stimulation during homeostatic behaviors. PMID:16275919

Wideband feeds have developed for Kashima 34m antenna and new 2.4m portable VLBI antennas. Prototypes of the wideband feeds are multimode horns, first one was set on 34m in the end of 2013, and then replaced next one with 6.5-15.0GHz receiving frequency. Now, a new feed for 3.2GHz-14.4GHz will be installed in 2.4m and 34m antennas in this spring, which are named NINJA feed, because of its design flexibility in beam shpae. Next, IGUANA feed is now under design and fabrication, which is aimed for 2.2-22GHz and covers VGOS(VLBI2010) specification. This has coaxial structure, the smaller "daughter feed" for 6.4-22GHz is placed in the center of the larger "Mother feed" for 2.2-6.4GHz.They are used for our project of time and frequency transfer between remote atomic clocks by wideband VLBI, named Gala-V(Garapagos VLBI), and will also be used wideband VLBI observation for astronmy and geodesy.Prototype feeds were tested in measurement of aperture efficiency, SEFD and Tsys of 34m "Super Kashima Antenna" and both 6.7/12.2GHz methanol maser detection in one reciever system, and then better one is used for wideband VLBI observations.

In cases in which expressed breast milk is given or breast feeding cannot be done at all, a suitable alternative to breast feeding is still in question. Bottle feeding poses many hazards. Spoon and bowl have been proposed as a reasonable alternative since users can achieve better cleanliness. Yet it is impractical for staff in health facility nurseries to feed every newborn with the spoon and bowl method since it requires so much time. On average, they need to provide oral feeds to at least 10 babies a day. In India, maternal grandparents present the family of a newborn with the traditional Sindhi silver or stainless steel utensil to provide the infant drinking water. It is called Suthi. It holds either 10 or 20 cc, making it easier to quantify the amount of milk/feed. It has a long semicircular beak and curved rounded margins. Advantages of the Suthi over other alternative feeding methods include: it is a shallow container with a broad upper surface, allowing the user to clean it thoroughly and easily; its narrow beak can go directly into the mouth of the newborn, particularly premature infants and low birth weight infants, with relative ease, reducing the likelihood of spilling milk, and the Suthi feeding procedure is less messy and faster (7-10 vs. 15-20 minutes for spoon) than other procedures. The Suthi method allows nursery staff to spend less time per feed for 8-10 babies (minimum time needed, 1 vs. 3 hours). It can also be used at home. Its use will reduce the likelihood of aspiration since it cannot be used lying down. Since it is a traditional container, the community will accept Suthi. When breast feeding is not possible, the Suthi should be used.

The term feed efficiency is vague, and is defined differently by people. Historically, feed efficiency has been defined as the feed:gain (F:G) ratio or the inverse (G:F). Indexes have been developed to rank animals for feed efficiency. These indexes include residual feed intake (RFI) and residual...

The fall armyworm Spodoptera frugiperda is an economically important pest of small grain crops that occurs in all maize growing regions of the Americas. The intensive use of chemical pesticides for its control has led to the selection of resistant populations, however, to date, the molecular mechanisms underlying resistance have not been characterised. In this study the mechanisms involved in the resistance of two S. frugiperda strains collected in Brazil to chlorpyrifos (OP strain) or lambda-cyhalothrin (PYR strain) were investigated using molecular and genomic approaches. To examine the possible role of target-site insensitivity the genes encoding the organophosphate (acetylcholinesterase, AChE) and pyrethroid (voltage-gated sodium channel, VGSC) target-site proteins were PCR amplified. Sequencing of the S. frugiperda ace-1 gene identified several nucleotide changes in the OP strain when compared to a susceptible reference strain (SUS). These result in three amino acid substitutions, A201S, G227A and F290V, that have all been shown previously to confer organophosphate resistance in several other insect species. Sequencing of the gene encoding the VGSC in the PYR strain, identified mutations that result in three amino acid substitutions, T929I, L932F and L1014F, all of which have been shown previously to confer knockdown/super knockdown-type resistance in several arthropod species. To investigate the possible role of metabolic detoxification in the resistant phenotype of the OP and PYR stains all EST sequences available for S. frugiperda were used to design a gene-expression microarray. This was then used to compare gene expression in the resistant strains with the susceptible reference strain. Members of several gene families, previously implicated in metabolic resistance in other insects were found to be overexpressed in the resistant strains including glutathione S-transferases, cytochrome P450s and carboxylesterases. Taken together these results provide

Exposure to Bacillus thuringiensis (Bt) toxins in low- and moderate-dose transgenic crops may induce sublethal effects and increase the rate of Bt resistance evolution, potentially compromising control efficacy against target pests. We tested this hypothesis using the fall armyworm Spodoptera frugiperda, a major polyphagous lepidopteran pest relatively tolerant to Bt notorious for evolving field-relevant resistance to single-gene Bt maize. Late-instar larvae were collected from Bt Cry1Ab and non-Bt maize fields in five locations in Brazil, and their offspring was compared for survival, development, and population growth in rearing environment without and with Cry1Ab throughout larval development. Larval survival on Cry1Ab maize leaves varied from 20 to 80% among the populations. Larvae reared on Cry1Ab maize had seven-day delay in development time in relation to control larvae, and such delay was shorter in offspring of armyworms from Cry1Ab maize. Population growth rates were 50–70% lower for insects continuously exposed to Cry1Ab maize relative to controls, showing the population-level effect of Cry1Ab, which varied among the populations and prior exposure to Cry1Ab maize in the field. In three out of five populations, armyworms derived from Bt maize reared on Cry1Ab maize showed higher larval weight, faster larval development and better reproductive performance than the armyworms derived from non-Bt maize, and one of these populations showed better performance on both Cry1Ab and control diets, indicating no fitness cost of the resistance trait. Altogether, these results indicate that offspring of armyworms that developed on field-grown, single-gene Bt Cry1Ab maize had reduced performance on Cry1Ab maize foliage in two populations studied, but in other three populations, these offspring had better overall performance on the Bt maize foliage than that of the armyworms from non-Bt maize fields, possibly because of Cry1Ab resistance alleles in these populations

The application of silica nanoparticles (NPs) in the biomedical field experienced a great development. The driving forces for these and future developments are the possibility to design NPs with homogeneous size and structure amenable to specific grafting. Moreover, it is possible to tune the characteristics of the NPs to meet the requirements of each specific cell and desired application. Herein, we analyzed the effect of silica NPs of various sizes and surface charge on the viability of Spodoptera frugiperda cells (Sf9 cell line) with the aim of extending the knowledge of possible toxicity of the NPs in the environment and development of new tools for insect control. Moreover, these results will also contribute to develop more effective systems for gene vectors delivery and recombinant proteins expression. Bare silica NPs of 14 nm, 380 nm and 1430 nm as well as amine-modified silica NPs of 131 nm and 448 nm were obtained by the Stöber method. The NPs were characterized by DLS and zeta potential measurements. The cell viability was assessed by the MTT test. It was observed that the 14 nm NPs possess the highest toxic effect. Indeed, after 24 h, the viability of the cells exposed to the lower concentration of NPs (0.12 mg/ml) was about 40% of the value obtained for the control cells not exposed to NPs. Moreover, the exposure to other negative charged NPs also causes a lower activity when compared with the control. Alternatively, lower concentrations of positive charged NPs (i.e.: 0.12 or 0.6 mg/ml) demonstrated to stimulate the proliferation of the cells and higher concentrations (i.e.: 7.2 mg/ml) did not present significant differences with the control. In conclusion, we have demonstrated that the NPs possess an effect that is highly influenced by the size, charge and concentration. Although, silica NPs are being used in the biomedical field, these results contribute to further understanding the risk that could be associated to nanoparticles and how these can be

Terpenoids, which have many biological activities and have occurred widely in nature, can be artificially synthesized. However, regioselective oxidation of terpenoids is difficult by chemical methods. In this study, (+)- and (-)-citronellene were biotransformed with Spodoptera litura to define the mechanism of metabolism of citronellene and gain a new natural terpenoid. (+)-Citronellene was converted to (2S,3S)-3,7-dimethyl-6-octene-1,2-diol and (2R,3S)-3,7-dimethyl-6-octene-1,2-diol (89.7%), (3S,6S)-(-)-3,7-dimethyl-1-octene-6,7-diol (3.8%), (3S)-(6E)-(+)-3,7-dimethyl-1,6-octadien-8-ol (4.2%), and (3S)-(6E)-(+)-3,7-dimethyl-1,6-octadien-8-oic acid (2.3%). In contrast, (-)-citronellene was converted to (2R,3R)-3,7-dimethyl-6-octene-1,2-diol and (2S,3R)-3,7-dimethyl-6-octene-1,2-diol (56.3%), (+)-iridan-7,8-diol (3.5%), and (3R)-(6E)-(-)-3,7-dimethyl-1,6-octadien-8-oic acid (40.2%). The main metabolic pathway of (+)- and (-)-citronellene by larvae of S. litura was oxidized at the terminal double bond and trans-allylic methyl position. Particularly on (+)-citronellene, the regioselective reaction was shown. On the oxidation of C-6, C-7, and C-8 positions, four new compounds (3S,6S)-(-)-3,7-dimethyl-1-octene-6,7-diol, (3S)-(6E)-(+)-3,7-dimethyl-1,6-octadien-8-oic acid, (+)-iridan-7,8-diol, and (3R)-(6E)-(-)-3,7-dimethyl-1,6-octadien-8-oic acid were produced in regioselective oxidation. It noted that stereoselective oxidation occurred between the enantiomers. The C-6 position was oxidized on the (+)-(3S) form, whereas cyclized and the C-7 position were oxidized on the (-)-(3R) form.

Drosophila mojavensis wrigleyi and D. mojavensis sonorensis are geographically separated races of cactophilic fruit flies. D. mojavensis sonorensis inhabits the Sonoran Desert and utilizes necrotic rots of Stenocereus thurberi Engelm. as a food source and to oviposit while D. mojavensis wrigleyi inhabits Santa Catalina Island, California and utilizes the necrotic rots of Opuntia littoralis (Engelm.) Cockerell. The objectives of this study were to determine the volatile compositions of the necrotic cacti and to determine if the volatile components show either selective toxicity or attraction toward the fruit flies. The volatile chemical compositions of field-rot specimens of both necrotic cacti were obtained by dynamic headspace (purge-and-trap) and hydrodistillation techniques and analyzed by gas chromatography - mass spectrometry. The volatile fraction of necrotic S. thurberi early rot was dominated by carboxylic acids (84.8%) and the late rot by p-cresol (32.6% in the dynamic headspace sample and 55.9% in the hydrodistilled sample). O. littoralis volatiles were dominated by carboxylic acids (86% in the dynamic headspace sample and 89.1% in the hydrodistilled sample). Fifteen compounds that were identified in the necrotic rot volatiles were used to test insecticidal activity and olfactory preference on the cactophilic Drosophila species, as well as D. melanogaster. Differences in toxicity and olfactory preference were observed between the different taxa. Both races of D. mojavensis exhibited toxicity to benzaldehyde and 2-nonanone, while butanoic acid and palmitic acid were tolerated at high concentrations. D. m. wrigleyi demonstrated a greater olfactory preference for anisole, butanoic acid, 2-heptanone, and palmitic acid than did D. m. sonorensis, while D. m. sonorensis demonstrated a greater preference for hexadecane, octanoic acid, and oleic acid than did D. m. wrigleyi.

This is a report of an in vitro study evaluating the ability of six solutions to dissolve clotted enteral feeding, which can cause feeding tube occlusion. The following clotted enteral feeding products were tested: Ensure Plus, Ensure Plus with added protein (Promod 20 g/liter), Osmolite, Enrich, and Pulmocare. Clot dissolution was then tested by adding Adolf's Meat Tenderizer, Viokase, Sprite, Pepsi, Coke, or Mountain Dew. Distilled water served as control. Dissolution score for each mixture was assessed blindly. Best dissolution was observed with Viokase in pH 7.9 solution (p less than 0.01). Similar results were obtained when feeding tube patency was restored in eight in vitro occluded feeding tubes (Dobbhoff, French size 8) by using first Pepsi (two/eight successful) and then Viokase in pH 7.9 (six/six successful). We also report our experience in the first 10 patients with occluded feeding tubes using this Viokase solution injected through a Drum catheter into the feeding tube. In seven patients, this method proved to be successful, and the reasons for failure in three patients include a knotted tube, impacted tablet powder, and a formula clot fo 24 hr duration and 45 cm in length.

Laboratory work was completed on a set of evaporation tests designed to establish a feed envelope for the fractional crystallization process. The feed envelope defines chemical concentration limits within which the process can be operated successfully. All 38 runs in the half-factorial design matrix were completed successfully, based on the qualitative definition of success. There is no feed composition likely to be derived from saltcake dissolution that would cause the fractional crystallization process to not meet acceptable performance requirements. However, some compositions clearly would provide more successful operation than other compositions.

Neuromodulators play a key role in adjusting animal behavior based on environmental cues and internal needs. Here, we review the regulation of Drosophila feeding behavior to illustrate how neuromodulators achieve behavioral plasticity. Recent studies have made rapid progress in determining molecular and cellular mechanisms that translate the metabolic needs of the fly into changes in neuroendocrine and neuromodulatory states. These neuromodulators in turn promote or inhibit discrete feeding behavioral subprograms. This review highlights the links between physiological needs, neuromodulatory states, and feeding decisions. PMID:24937262

A coal feed lock is provided for dispensing coal to a high pressure gas producer with nominal loss of high pressure gas. The coal feed lock comprises a rotor member with a diametral bore therethrough. A hydraulically activated piston is slidably mounted in the bore. With the feed lock in a charging position, coal is delivered to the bore and then the rotor member is rotated to a discharging position so as to communicate with the gas producer. The piston pushes the coal into the gas producer. The rotor member is then rotated to the charging position to receive the next load of coal.

Essential oils from Greek Mentha species showed different chemical compositions for two populations of M. pulegium, characterized by piperitone and pulegone. Mentha spicata essential oil was characterized by endocyclic piperitenone epoxide, piperitone epoxide, and carvone. The bioactivities of these essential oils and their components have been tested against insect pests (Leptinotarsa decemlineata, Spodopteralittoralis and Myzus persicae), root-knot nematodes (Meloydogine javanica) and plants (Lactuca sativa, Lolium perenne, Solanum lycopersicum). The structure-activity relationships of these compounds have been studied including semi-synthetic endocyclic trans-carvone epoxide, exocyclic carvone epoxide, a new exocyclic piperitenone epoxide and trans-pulegone epoxide. Leptinotarsa decemlineata feeding was affected by piperitenone and piperitone epoxide. Spodopteralittoralis was affected by piperitone epoxide and pulegone. The strongest nematicidal agent was piperitenone epoxide, followed by piperitone epoxide, piperitenone and carvone. Germination of S. lycopersicum and L. perenne was significantly affected by piperitenone epoxide. This compound and carvone epoxide inhibited L. perenne root and leaf growth. Piperitenone epoxide also inhibited the root growth of S. lycopersicum. The presence of a C(1) epoxide resulted in strong antifeedant, nematicidal and phytotoxic compounds regardless of the C(4) substituent. New natural crop protectants could be developed through appropriate structural modifications in the p-menthane skeleton.

... you choose to breastfeed or formula feed. About Breastfeeding Breastfeeding your newborn has many advantages. Perhaps most ... to care for her newborn. continue Limitations of Breastfeeding With all the good things known about breastfeeding, ...

... this page: //medlineplus.gov/ency/patientinstructions/000803.htm Breastfeeding vs. Formula Feeding To use the sharing features ... best for you and your family. Benefits of Breastfeeding Breastfeeding is a wonderful way to bond with ...

Lectins are carbohydrate-interacting proteins that play a pivotal role in multiple physiological and developmental aspects of all organisms. They can specifically interact with different bacterial and viral pathogens through carbohydrate-recognition domains (CRD). In addition, lectins are also of biotechnological interest because of their potential use as biosensors for capturing and identifying bacterial species. In this work, three C-type lectins from the Lepidoptera Spodoptera exigua were produced as recombinant proteins and their bacterial agglutination properties were characterized. The lowest protein concentration producing bacterial agglutination against a panel of different Gram+ and Gram− as well as their carbohydrate binding specificities was determined for the three lectins. One of these lectins, BLL2, was able to agglutinate cells from a broad range of bacterial species at an extremely low concentration, becoming a very interesting protein to be used as a biosensor or for other biotechnological applications involving bacterial capture. PMID:28257054

The autumn return migration of the beet armyworm Spodoptera exigua Hübner and other insects in northern China was observed with radar and with a simultaneously-operated searchlight trap and ground light-trap at a site in Langfang, near Beijing. The searchlight trap was found to be an efficient tool for trapping migrants and, operated alongside a ground light-trap, could distinguish migrant from locally-flying species. It was confirmed that S. exigua and some other species were high-altitude nocturnal windborne migrants during September and October in northern China. Maximum density of moths typically occurred below 500 m, and strong layering was often observed at about 200 m above ground level in airflows that would carry the moths towards the south. Descent of S. exigua in the vicinity of the radar site in late September was often associated with rain.

Lectins are carbohydrate-interacting proteins that play a pivotal role in multiple physiological and developmental aspects of all organisms. They can specifically interact with different bacterial and viral pathogens through carbohydrate-recognition domains (CRD). In addition, lectins are also of biotechnological interest because of their potential use as biosensors for capturing and identifying bacterial species. In this work, three C-type lectins from the Lepidoptera Spodoptera exigua were produced as recombinant proteins and their bacterial agglutination properties were characterized. The lowest protein concentration producing bacterial agglutination against a panel of different Gram+ and Gram- as well as their carbohydrate binding specificities was determined for the three lectins. One of these lectins, BLL2, was able to agglutinate cells from a broad range of bacterial species at an extremely low concentration, becoming a very interesting protein to be used as a biosensor or for other biotechnological applications involving bacterial capture.

Field-evolved resistance to maize event TC1507 expressing the Cry1Fa toxin from Bacillus thuringiensis (Bt) was detected in populations of Spodoptera frugiperda from Puerto Rico. We tested for cross-resistance to purified Cry1A toxins and commercial Bt pesticides in susceptible (Benzon) and TC1507-resistant (456) strains of S. frugiperda. Larvae from the 456 strain exhibited cross-resistance to Cry1Ab and Cry1Ac toxins, while no differences in susceptibility to XenTari WG and DiPel ES pesticides were detected. These data support cross-resistance to toxins that share binding sites with Cry1Fa and no cross-resistance to Bt pesticides in S. frugiperda with field-evolved resistance to Bt maize.

The development of transgenic maize plants expressing soybean proteinase inhibitors could reduce the economic damage of one of the major maize pests in Brazil, the fall armyworm, Spodoptera frugiperda (J.E. Smith, 1797). We examined the influence of soybean proteinase inhibitors on digestive enzyme properties and development of S. frugiperda larvae. The inhibition of trypsin and chymotrypsin activities in vitro by soybean proteinase inhibitors suggested that either Kunitz (SBTI) or Bowman-Birk (SBBI) would have a potential antimetabolic effect when ingested by insect larvae. However, chronic ingestion of semipurified soybean inhibitors did not result in a significant reduction of growth and development of fall armyworm. Therefore, digestive serine proteinase activities (trypsin and chymotrypsin) of fall armyworm larvae were characterized. The results suggest that S. frugiperda was able to physiologically adapt to dietary proteinase inhibitors by altering the complement of proteolytic enzymes in the insect midguts.

Nowadays molecular species delimitation methods promote the identification of species boundaries within complex taxonomic groups by adopting innovative species concepts and theories (e.g. branching patterns, coalescence). As some of them can efficiently deal with large single-locus datasets, they could speed up the process of species discovery compared to more time consuming molecular methods, and benefit from the existence of large public datasets; these methods can also particularly favour scientific research and actions dealing with threatened or economically important taxa. In this study we aim to investigate and clarify the status of economically important moths species belonging to the genus Spodoptera (Lepidoptera, Noctuidae), a complex group in which previous phylogenetic analyses and integrative approaches already suggested the possible occurrence of cryptic species and taxonomic ambiguities. In this work, the effectiveness of innovative (and faster) species delimitation approaches to infer putative species boundaries has been successfully tested in Spodoptera, by processing the most comprehensive dataset (in terms of number of species and specimens) ever achieved; results are congruent and reliable, irrespective of the set of parameters and phylogenetic models applied. Our analyses confirm the existence of three potential new species clusters (for S. exigua (Hübner, 1808), S. frugiperda (J.E. Smith, 1797) and S. mauritia (Boisduval, 1833)) and support the synonymy of S. marima (Schaus, 1904) with S. ornithogalli (Guenée, 1852). They also highlight the ambiguity of the status of S. cosmiodes (Walker, 1858) and S. descoinsi Lalanne-Cassou & Silvain, 1994. This case study highlights the interest of molecular species delimitation methods as valuable tools for species discovery and to emphasize taxonomic ambiguities. PMID:25853412

Highlights: Black-Right-Pointing-Pointer Baculovirus p35 is regulated by both viral and host factors. Black-Right-Pointing-Pointer Baculovirus p35 is negatively regulated by SfP53-like factor. Black-Right-Pointing-Pointer Baculovirus p35 is positively regulated by SfAP-1-like factor. -- Abstract: Baculovirus p35 belongs to the early class of genes of AcMNPV and requires viral factors like Immediate Early protein-1 for its transcription. To investigate the role of host factors in regulating p35 gene expression, the putative transcription factor binding sites were examined in silico and the role of these factors in influencing the transcription of p35 gene was assessed. We focused our studies on AP-1 and P53-like factors, which are activated under oxidative stress conditions. The AP-1 motif is located at -1401 while P53 motif is at -1912 relative to p35 translation start site. The predicted AP-1 and P53 elements formed specific complexes with Spodoptera frugiperda nuclear extracts. Both AP-1 and P53 motif binding proteins were down regulated as a function of AcMNPV infection in Spodoptera cells. To address the question whether during an oxidative outburst, the p35 transcription is enhanced; we investigated the role of these oxidative stress induced host transcription factors in influencing p35 gene transcription. Reporter assays revealed that AP-1 element enhances the transcription of p35 by a factor of two. Interestingly, P53 element appears to repress the transcription of p35 gene.

Spodoptera frugiperda (Smith 1797) (Lepidoptera: Noctuidae) is a major pest of maize, Zea mays L. Its control is often achieved through repeated applications per season of insecticides, which may lead to adverse effects on the ecosystem. Thus, the study of alternative methods with less environmental impact has expanded to include the use of essential oils. These oils are products of the secondary metabolism in plants, and their insecticidal activity has been widely demonstrated in populations of many pest insects. This study evaluated the insecticidal activities of essential oils from Eucalyptus staigeriana, Ocimum gratissimum, and Foeniculum vulgare on Spodoptera frugiperda. Gas chromatography–mass spectrometry profiles and contact toxicity of these oils as well as their sublethal effects on larvae and reproductive parameters in adults were evaluated. All three oils had sublethal effects on S. frugiperda; however, the oil of O. gratissimum showed the best results at all doses tested. These essential oils may have promise for control of S. frugiperda.

Tube feeding is an integral part of medical therapies, and can be easily managed also in the outpatient setting. Tube feeding by the stomach or small intestine with nasogastral or nasojejunal tubes is common in clinical practice. Long-term nutrition is usually provided through a permanent tube, i. e. a percutaneous endoscopic gastrostomy (PEG). Modern portable nutrition pumps are used to cover the patient's nutritional needs. Enteral nutrition is always indicated if patients can not or should not eat or if nutritional requirements cannot be covered within 3 days after an intervention, e. g. after abdominal surgery. Industrially produced tube feedings with defined substrate concentrations are being used; different compositions of nutrients, such as glutamine fish oil etc., are used dependent on the the condition of the patient. Enteral nutrition may be associated with complications of the tube, e. g. dislocation, malposition or obstruction, as well as the feeding itself, e. g.hyperglycaemia, electrolyte disturbances, refeeding syndrome diarrhea or aspiration). However, the benefit of tube feeding usually exceeds the potential harm substantially.

Key impediments to increased corn yield and quality in the southeastern US coastal plain region are damage by ear-feeding insects and aflatoxin contamination caused by infection of Aspergillus flavus. Key ear-feeding insects are corn earworm, Helicoverpa zea, fall armyworm, Spodoptera frugiperda, maize weevil, Sitophilus zeamais, and brown stink bug, Euschistus servus. In 2006 and 2007, aflatoxin contamination and insect damage were sampled before harvest in three 0.4-hectare corn fields using a grid sampling method. The feeding damage by each of ear/kernel-feeding insects (i.e., corn earworm/fall armyworm damage on the silk/cob, and discoloration of corn kernels by stink bugs), and maize weevil population were assessed at each grid point with five ears. The spatial distribution pattern of aflatoxin contamination was also assessed using the corn samples collected at each sampling point. Aflatoxin level was correlated to the number of maize weevils and stink bug-discolored kernels, but not closely correlated to either husk coverage or corn earworm damage. Contour maps of the maize weevil populations, stink bug-damaged kernels, and aflatoxin levels exhibited an aggregated distribution pattern with a strong edge effect on all three parameters. The separation of silk- and cob-feeding insects from kernel-feeding insects, as well as chewing (i.e., the corn earworm and maize weevil) and piercing-sucking insects (i.e., the stink bugs) and their damage in relation to aflatoxin accumulation is economically important. Both theoretic and applied ramifications of this study were discussed by proposing a hypothesis on the underlying mechanisms of the aggregated distribution patterns and strong edge effect of insect damage and aflatoxin contamination, and by discussing possible management tactics for aflatoxin reduction by proper management of kernel-feeding insects. Future directions on basic and applied research related to aflatoxin contamination are also discussed.

The invention is a dust feed device for delivery of a uniform supply of dust for long periods of time to an aerosolizing means for production of a dust suspension. The device utilizes at least two tandem containers having spiral brushes within the containers which transport the dust from a supply to the aerosolizer means.

Over the past several years, a number of randomized controlled trials have compared the effects of breastfeeding and formula feeding and the effects of docosahexaenoic acid (DHA)–supplemented and non-supplemented formulas on visual function in both preterm and term infants. Some studies have shown b...

Globally, aquaculture has been growing at a rapid pace (currently 8.5% per year) over the past two decades, and is recognized as the fastest growing food production sector of agriculture in the U.S. Growth of aquaculture and other industries (e.g., other monogastric and ruminant livestock feed appl...

Language arts teachers and library media specialists bear the responsibility of teaching students how to properly feed their brains. In this article, the author describes how she teaches her students to make wise choices when selecting books. Furthermore, she presents the "Brain Food Pyramid" model that looks similar to the food pyramid but it…

The feeding of Diarmis proboscis is an exciting outdoor laboratory activity that demonstrates a single concept of adaptations--cryptic colorations. The students are "transformed" into D. proboscis (no Harry Potter magic needed) in order to learn how adaptations work in the natural world. Prior to beginning this activity, students should have a…

The objectives of this study were to determine the effect of feed presentation on meal frequency and duration, as well as diurnal feeding patterns of dairy calves, and to assess any longer-term differences in feeding patterns resulting from previous experience. Twenty Holstein bull calves were exposed from wk 1 to 8 of life to 1 of 2 feed presentation treatments: concentrate and chopped grass hay (<2.5 cm) offered at a ratio of 7:3 as a mixture (MIX), or as separate components (COM). Feed was provided ad libitum. Calves received 8L/d of milk replacer (1.2 kg of dry matter), with the amount progressively reduced after 5 wk to facilitate weaning by the end of wk 7. At the beginning of wk 9, all calves received the MIX diet and remained on trial for an additional 3 wk. Feeding behavior was recorded from video for 4d during wk 6, 8, 9, and 11. In wk 6, calves fed MIX spent more time feeding than calves fed COM (56.7 vs. 46.8 min/d). In wk 8, calves fed MIX spent more time feeding (174.0 vs. 139.1 min/d) and had a lower rate of intake (11.5 vs. 14.7 g/min) compared with calves fed COM. Meal frequency was similar between treatments (12.2 meals/d). Diurnal feeding patterns in wk 8 were also affected by feed presentation, with calves fed MIX spending less time feeding at time of feed delivery and more time feeding throughout the rest of the daylight hours than calves fed COM. Diurnal feeding patterns of hay and concentrate in wk 8 differed for calves fed COM, with more time spent consuming hay at time of feed delivery and less time spent consuming hay throughout the rest of the day. Once calves previously fed COM were transitioned to the MIX diet in wk 9, meal frequency, meal duration, and diurnal feeding patterns were similar between treatments: both treatments spent similar amounts of time feeding (173.9 min/d) and had similar peaks in feeding activity at time of feed delivery, sunrise, and sunset. Provision of hay and concentrate to young calves as a mixed ration

Animals depend on reliable sensory information for accurate behavioural decisions. For herbivorous insects it is crucial to find host plants for feeding and reproduction, and these insects must be able to differentiate suitable from unsuitable plants. Volatiles are important cues for insect herbivores to assess host plant quality. It has previously been shown that female moths of the Egyptian cotton leafworm, Spodopteralittoralis (Lepidoptera: Noctuidae), avoid oviposition on damaged cotton Gossypium hirsutum, which may mediated by herbivore-induced plant volatiles (HIPVs). Among the HIPVs, some volatiles are released following any type of damage while others are synthesized de novo and released by the plants only in response to herbivore damage. In behavioural experiments we here show that oviposition by S. littoralis on undamaged cotton plants was reduced by adding volatiles collected from plants with ongoing herbivory. Gas chromatography-electroantennographic detection (GC-EAD) recordings revealed that antennae of mated S. littoralis females responded to 18 compounds from a collection of headspace volatiles of damaged cotton plants. Among these compounds, a blend of the seven de novo synthesized volatile compounds was found to reduce oviposition in S. littoralis on undamaged plants under both laboratory and ambient (field) conditions in Egypt. Volatile compounds that are not produced de novo by the plants did not affect oviposition. Our results show that ovipositing females respond specifically to the de novo synthesized volatiles released from plants under herbivore attack. We suggest that these volatiles provide reliable cues for ovipositing females to detect plants that could provide reduced quality food for their offspring and an increased risk of competition and predation.

In this review I outline studies on wild non-human primates using information on feeding rate, which is defined as the food intake per minute on a dry-weight basis; further, I summarize the significance of feeding rate in primate feeding ecology. The optimal foraging theory has addressed three aspects of animal feeding: (1) optimal food patch choice, (2) optimal time allocation to different patches, and (3) optimal food choice. In order to gain a better understanding of these three aspects, the feeding rate itself or its relevance indices (e.g., rates of calorie and protein intake) could be appropriate measures to assess the quality of food and food patches. Moreover, the feeding rate plays an essential role in estimation of total food intake, because it varies greatly for different food items and the feeding time is not a precise measure. The feeding rate could also vary across individuals who simultaneously feed on the same food items in the same food patch. Body size-dependent and rank-dependent differences in the feeding rate sometimes cause individuals to take strategic behavioral options. In the closing remarks, I discuss the usefulness of even limited data on feeding rate obtained under adverse observational conditions in understanding primate feeding ecology.

Few physicians in the United States receive formal education related to principles of infant care in Islamic families. Breastfeeding has a religious basis in Islam and it is recommended that the mother suckle her offspring for 2 years if possible. Weaning from the breast before that period is allowed if mutually decided on by both parents. The infant's father has an obligation to support his wife through any circumstances that may affect breastfeeding and, in case of divorce, provide shelter and financial support to the mother-infant dyad for as long as breastfeeding continues. By showing understanding and respect of Islamic beliefs related to infant feeding, clinicians can help support healthy early feeding of Muslim infants.

Peritrophic membrane proteins are important components of the insect peritrophic membrane. A novel cDNA gene encoding a chitin-binding protein, named secbp66, was identified by immunization screening of the cDNA library of Spodoptera exigua. The full length of secbp66 is 1806 bp, which encodes 602 amino acids. The predicted weight of the protein is 64.2 kDa. Bioinformatic analysis showed that a signal peptide composed of 17 amino acids located at the N-terminal of SeCBP66 contained seven tandem putative Type-II functional chitin-binding domains and five potential N-glycosylation sites, but no O-linked glycosylation sites. To study the properties of SeCBP66, recombinant SeCBP66 was successfully expressed in the insect cell line BTI-Tn-5B1-4 with a Bac-to-Bac expression system. A chitin binding experiment showed that the recombinant SeCBP66 protein could bind to chitin strongly. This study of the novel chitin-binding protein SeCBP66 provides a basis for developing new control targets for S. exigua.

The present studies were carried out to evaluate resistance in the populations of Spodoptera litura Fab. (Lepidoptera, Noctuidae) from five districts of Hunan Province in China to various insecticides from 2010 to 2012 using a standard leaf dip bioassay method. For organophosphates and pyrethroids, resistance ratios compared with a susceptible Lab-BJ strain were in the range of 14-229-fold for organophosphates and 12-227-fold for pyrethroids. Similarly, relative low levels of resistance to emamectin, indoxacarb, and chlorfenapyr were observed in all five populations. In contrast, the resistance to carbamates (thiodicarb or methomyl) was significantly higher than that of organophosphates, pyrethroids and newer chemistry insecticides. The pairwise correlation coefficients of LC50 values indicated that the newer chemistry insecticides and old generation insecticides were not significant except abamectin, which was negatively significantly correlated with methomyl. A significant correlation was observed between thiodicarb, methomyl, and deltamethrin, whereas resistance to bifenthrin showed no correlations with resistance to other insecticides except deltamethrin. The results are discussed in relation to integrated pest management for S. litura with special reference to management of field evolved resistance to insecticides.

To examine the responses of the secondary lepidopteran pest Spodoptera litura to two Bacillus thuringiensis (Bt) corn hybrids [5422Bt1 (Event Bt11), 5422CBCL (MON810)] expressing Cry1Ab, larval bioassays with Cry1Ab toxin, corn leaves or kernels and bagging on corn plants were conducted. The results showed that larvae displayed a similar performance when fed kernels, but not leaves of 5422Bt1, 5422CBCL and their near-isogenic non-Bt corn (5422). Significantly higher Cry1Ab amounts were detected in larvae fed leaves than kernels of both Bt hybrids, with different molecular weights of protein band in plants (72 and 90 kDa for 5422Bt1 and 5422CBCL, respectively), gut contents (65 kDa), feces (50 kDa), which indicated that larvae had lower ingestion, higher degradation and excretion of Cry1Ab when fed kernels not leaves of both Bt hybrids. Significantly higher levels of cadherin-like receptors and alkaline phosphatase transcripts were detected in larvae fed leaves than kernels of two Bt hybrids. Catalase, superoxide dismutase and glutathione-S-transferase activities in larvae fed 5422Bt1 leaves were significantly higher than that of 5422 treatments. Therefore, S. litura had low susceptibility to 5422Bt1 and 5422CBCL when larvae fed kernels not leaves of Bt corn. Additionally, S. litura presented a much stronger tolerance to 5422CBCL than 5422Bt1.

Genetically modified cotton that produces a crystalline protein from Bacillus thuringiensis subsp. kurstaki (Berliner) (Bt) has been widely deployed to manage lepidopteran insect pests in cotton growing areas worldwide. However, susceptibility of different insect species to Bt protein varies, which may affect lepidopteran pest populations in the field. Studies on effects of two transgenic cotton lines (BG1560 and GK19) carrying a Cry1A gene on common cutworm Spodoptera litura F. (Lepidoptera: Noctuidae), were conducted during 2002-2005 in the cotton planting region of the Yangtze River valley of China. Results showed that common cutworm larvae had low susceptibility to Bt cotton. There was no significant difference in larval population densities in conventional and Bt cotton fields. However, the larval populations of the insect on conventional plants treated with chemical insecticides for control of target pest of Bt cotton were significantly lower than that in Bt cotton fields. These results indicated that the common cutworm was the potential to become a major and alarming pest in Bt cotton fields, and therefore efforts to develop an effective alternative management strategy are needed.

The armyworm, Spodoptera litura (F.) is a polyphagous and important agricultural pest worldwide. In this study, we examined the effect of electron beam irradiation on developmental stages, reproduction, and DNA damage of S. litura. Eggs (0-24 h old), larvae (3rd instar), pupae (3 days old after pupation), and adults (24 h after emergence) were irradiated with electron beam irradiation of six levels between 30 and 250 Gy. When eggs were irradiated with 100 Gy, egg hatching was completely inhibited. When the larvae were irradiated, the larval period was significantly delayed, depending on the doses applied. At 150 Gy, the fecundity of adults that developed from irradiated pupae was entirely inhibited. However, electron beam irradiation did not induce the instantaneous death of S. litura adults. Reciprocal crosses between irradiated and unirradiated moths demonstrated that females were more radiosensitive than males. We also conducted the comet assay immediately after irradiation and over the following 5 days period. Severe DNA fragmentation in S. litura cells was observed just after irradiation and the damage was repaired during the post-irradiation period in a time-dependent manner. However, at more than 100 Gy, DNA damage was not fully recovered.

Transgenic crop “pyramids” producing two or more Bacillus thuringiensis (Bt) toxins active against the same pest are used to delay evolution of resistance in insect pest populations. Laboratory and greenhouse experiments were performed with fall armyworm, Spodoptera frugiperda, to characterize resistance to Bt maize producing Cry1A.105 and Cry2Ab and test some assumptions of the “pyramid” resistance management strategy. Selection of a field-derived strain of S. frugiperda already resistant to Cry1F maize with Cry1A.105 + Cry2Ab maize for ten generations produced resistance that allowed the larvae to colonize and complete the life cycle on these Bt maize plants. Greenhouse experiments revealed that the resistance was completely recessive (Dx = 0), incomplete, autosomal, and without maternal effects or cross-resistance to the Vip3Aa20 toxin produced in other Bt maize events. This profile of resistance supports some of the assumptions of the pyramid strategy for resistance management. However, laboratory experiments with purified Bt toxin and plant leaf tissue showed that resistance to Cry1A.105 + Cry2Ab2 maize further increased resistance to Cry1Fa, which indicates that populations of fall armyworm have high potential for developing resistance to some currently available pyramided maize used against this pest, especially where resistance to Cry1Fa was reported in the field. PMID:26675246

Recently, wide attention has been given to the potential of recombinant baculovirus as a gene transfer vehicle for mammalian gene therapy. In this study, we packaged the recombinant baculoviruses with cytomegalovirus immediate-early (CMV-IE) promoter in Spodoptera frugiperda (Sf9) insect cells, and found that the CMV-IE promoter could efficiently drive the exogenic gene expression in the cells 12 h post-infection (h.p.i.). The expression level at 72 h.p.i. was only around half of that driven by polyhedrin promoter (Ppolh). However, the biological activity of the reporter proteins at 72 h.p.i. were similar with that driven by Ppolh. In addition, the Sf9 cells transfected with CMV-IE-containing plasmids also expressed foreign genes, suggesting that the CMV-IE-directed heterogeneous gene expression in the Sf9 cells was baculovirus-independent. These results demonstrate that the CMV-IE promoter might be used as a regular promoter in Sf9 cells.

Spinosad, a reduced-risk insecticide, acts on the nicotinic acetylcholine receptors and the gamma-aminobutyric acid receptor in the nervous system of target insects. However, its mechanism of action in non-neural insect cells is unclear. This study aimed to evaluate mitochondrial functional changes associated with spinosad in Spodoptera frugiperda (Sf9) insect cells. Our results indicate that in Sf9 cells, spinosad induces programmed cell death and mitochondrial dysfunction through enhanced reactive oxygen species production, mitochondrial permeability transition pore (mPTP) opening, and mitochondrial membrane potential collapse, eventually leading to cytochrome C release and apoptosis. The cytochrome C release induced by spinosad treatment was partly inhibited by the mPTP inhibitors cyclosporin A and bongkrekic acid. Subsequently, we found that spinosad downregulated Bcl-2 expression and upregulated p53 and Bax expressions, activated caspase-9 and caspase-3, and triggered PARP cleavage in Sf9 cells. These findings suggested that spinosad-induced programmed cell death was modulated by mitochondrial dysfunction and cytochrome C release.

Azadirachtin as a kind of botanical insecticide has been widely used in pest control. We previously reported that azadirachtin could induce apoptosis of Spodoptera litura cultured cell line Sl-1, which involves in the up-regulation of P53 protein. However, the detailed mechanism of azadirachtin-induced apoptosis is not clearly understood in insect cultured cells. The aim of the present study was to address the involvement of lysosome and lysosomal protease in azadirachtin-induced apoptosis in Sf9 cells. The result confirmed that azadirachtin indeed inhibited proliferation and induced apoptosis. The lysosomes were divided into different types as time-dependent manner, which suggested that changes of lysosomes were necessarily physiological processes in azadirachtin-induced apoptosis in Sf9 cells. Interestingly, we noticed that azadirachtin could trigger lysosomal membrane permeabilization and cathepsin L releasing to cytosol. Z-FF-FMK (a cathepsin L inhibitor), but not CA-074me (a cathepsin B inhibitor), could effectively hinder the apoptosis induced by azadirachtin in Sf9 cells. Meanwhile, the activity of caspase-3 could also be inactivated by the inhibition of cathepsin L enzymatic activity induced by Z-FF-FMK. Taken together, our findings suggest that azadirachtin could induce apoptosis in Sf9 cells in a lysosomal pathway, and cathepsin L plays a pro-apoptosis role in this process through releasing to cytosol and activating caspase-3.

Azadirachtin is one of the most effective botanical insecticides and has been widely used in pest control. Toxicological reports show that azadirachtin can induce apoptosis in various insect cell lines. However, studies of azadirachtin-induced autophagy in cultured insect cells are lacking. This study reports that azadirachtin A significantly inhibits cell proliferation by inducing autophagic and apoptotic cell death in Spodoptera litura cultured cell line (SL-1 cell). Characteristic autophagolysosome and Atg8-PE (phosphatidylethanolamine) accumulation were observed by electron microscopy and western blotting, indicating that azadirachtin triggered autophagy in SL-1 cell. Furthermore, azadirachtin inhibited survival signaling by blocking the activation of PI3K, AKT and the down-stream target of rapamycin. Similar to the positive control of starvation, azadirachtin induced the activation of insulin receptor (InR) via a cellular feedback mechanism. In addition, the autophagy-related 5 (Atg5), a molecular switch of autophagy and apoptosis, was truncated (tAtg5) to trigger cytochrome c release into the cytoplasm under azadirachtin stress, which indicated that azadirachtin induced apoptosis through autophagy. Our findings suggest that azadirachtin primarily induced autophagy in SL-1 cell by dysregulating InR- and PI3K/AKT/TOR pathways, then stimulated apoptosis by activating tAtg5. PMID:27752103

Research involving plants with insecticide activity evolved significantly in the last decades. Among these plants, the neem tree, Azadirachta indica, is commonly used against several insects, mainly Lepidoptera. The neem efficiency depends on the target insect and on the concentration used. A barrier against potential toxic agents ingested together with the food is the alimentary canal. Thus, this research aimed to describe the histology of the alimentary canal of Spodoptera frugiperda (J E Smith) larvae fed on leaves treated with neem (Neemseto) at a concentration of 0.5% and 1.0% and non treated, at different intervals (48, 96, 144, 192 and 240 h), by quantifying the regenerative cells and analyzing the secretion of the mesenteron histochemically. Larvae were immobilized at low temperatures (-4 degrees C), the alimentary canal was removed, fixed in Boüin s aqueous, embedded in paraplast and historesin, sectioned and stained with hematoxilin-eosin and periodic acid- Schiff. The histology of the alimentary canal of S. frugiperda was similar to other lepidopterans. Neem effects on morphology were seen only in the mesenteron, depending on the time and concentration used, such as: epithelium, reduction on regenerative cells and on the secretory activity in this region, confirmed by the histochemistry in both neem concentrations. These alterations were observed after 96 h at 1.0%, and 144 h at 0.5%. These results indicate that neem (Neemseto), at the concentrations studied, may be effective to control S. frugiperda because it promotes meaningful morphological alterations in the mesenteron.

The production of a multiple nucleopolyhedrovirus (SeMNPV) of the beet armyworm, Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae), has been markedly increased by using juvenile hormone analog (JHA) technology to generate a supernumerary sixth instar in the species. In the current study we compared the incidence of cannibalism in S. exigua fifth and sixth instars reared at low (two larvae per dish) and a high density (10 larvae per dish). The incidence of cannibalism was significantly higher in fifth instars compared with sixth instars and increased with rearing density on both instars. Infected larvae were more prone to become victims of cannibalism than healthy individuals in mixed groups comprising 50% healthy + 50% infected larvae in both instars reared at high density. Instar had a marked effect on occlusion body (OB) production because JHA-treated insects produced between 4.8- and 5.6-fold increase in OB production per dish compared with fifth instars at high and low densities, respectively. The insecticidal characteristics of OBs produced in JHA-treated insects, as indicated by LD50 values, were similar to those produced in untreated fourth or fifth instars. Because JHA technology did not increase the prevalence of cannibalism and had no adverse effect on the insecticidal properties of SeMNPV OBs, we conclude that the use of JHAs to generate a supernumerary instar is likely to be compatible with mass production systems that involve gregarious rearing of infected insects.

Fall armyworm, Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae), is a target species of transgenic corn (Zea mays L.) that expresses single and pyramided Bacillus thuringiensis (Bt) toxin. In 2014, S. frugiperda were collected from a light trap in North Carolina, and a total of 212 F1/F2 isofemale lines of S. frugiperda were screened for resistance to Bt and non-Bt corn. All of the 212 isolines were susceptible to corn tissue expressing Cry1A.105 + Cry2Ab, Cry1F + Cry1A.105 + Cry2Ab, and Cry1F + Cry1Ab + Vip3Aa20. Growth rate bioassays were performed to isolate non-recessive Bt resistance alleles. Seven individuals out of the 212 isofemale lines carried major non-recessive alleles conferring resistance to Cry1F. A pooled colony was created from the seven individuals. This colony was 151.21 times more resistant to Cry1F than a known-susceptible population and was also resistant to Cry1A.105, but was not resistant to Cry2Ab and Vip3Aa20. The results demonstrate that field populations of S. frugiperda collected from North Carolina are generally susceptible to Cry1F, but that some individuals carry resistant alleles. The data generated in this study can be used as baseline data for resistance monitoring. PMID:27119741

Vip3A is an 89-kDa protein secreted by Bacillus thuringiensis during vegetative growth. To determine the importance of Vip3A for the insect pathogenicity of B. thuringiensis the vip3A gene was deleted from strain HD1, yielding strain HD1Deltavip3A. Compared with HD1, strain HD1Deltavip3A was one-fourth as toxic to Agrotis ipsilon larvae and less than one-tenth as toxic to Spodoptera exigua larvae. When streptomycin was included in the S. exigua diet the toxicity of HD1Deltavip3A was approximately half that of HD1. Addition of HD1 spores increased the toxicity of purified Cry1 protein more than 600-fold against S. exigua, whereas addition of HD1Deltavip3A spores increased toxicity of Cry1 protein approximately 10-fold. These results demonstrate that an important component of B. thuringiensis insecticidal activity against S. exigua is the synthesis of Vip3A protein by B. thuringiensis cells after ingestion of spores and crystal proteins by insect larvae.

To examine the responses of the secondary lepidopteran pest Spodoptera litura to two Bacillus thuringiensis (Bt) corn hybrids [5422Bt1 (Event Bt11), 5422CBCL (MON810)] expressing Cry1Ab, larval bioassays with Cry1Ab toxin, corn leaves or kernels and bagging on corn plants were conducted. The results showed that larvae displayed a similar performance when fed kernels, but not leaves of 5422Bt1, 5422CBCL and their near-isogenic non-Bt corn (5422). Significantly higher Cry1Ab amounts were detected in larvae fed leaves than kernels of both Bt hybrids, with different molecular weights of protein band in plants (72 and 90 kDa for 5422Bt1 and 5422CBCL, respectively), gut contents (65 kDa), feces (50 kDa), which indicated that larvae had lower ingestion, higher degradation and excretion of Cry1Ab when fed kernels not leaves of both Bt hybrids. Significantly higher levels of cadherin-like receptors and alkaline phosphatase transcripts were detected in larvae fed leaves than kernels of two Bt hybrids. Catalase, superoxide dismutase and glutathione-S-transferase activities in larvae fed 5422Bt1 leaves were significantly higher than that of 5422 treatments. Therefore, S. litura had low susceptibility to 5422Bt1 and 5422CBCL when larvae fed kernels not leaves of Bt corn. Additionally, S. litura presented a much stronger tolerance to 5422CBCL than 5422Bt1. PMID:28186125

Biliverdin-binding vitellogenin (Vg) was purified from adult female hemolymph of the common cutworm, Spodoptera litura, by using gel filtration and ion exchange chromatographies. The molecular mass of the protein was 490 kDa and it was composed of two 188-kDa subunits. Three internal amino acid sequences obtained by digestion of the protein with lysylendopeptidase showed high similarity to those of Bombyx mori Vg, supporting the purified blue protein to be vitellogenin. latroscan analyses demonstrated the presence of biliverdin in Vg that occupied 2.4% of total lipid components. Among the lipids of Vg (9.5 micrograms total lipids per 100 micrograms protein), diacylglycerol was the most predominant, followed by phospholipid, hydrocarbons, and then triacylglycerol, while in biliverdin-binding proteins (BPs) purified from larval hemolymph (3.1 micrograms total lipids per 100 micrograms protein), phospholipid was the most abundant lipid followed by diacylglycerol; hydrocarbons and triacylglycerol were minor components. Vg was first detected in the hemolymph of female pupae one day before eclosion, but injection of 5 micrograms of methoprene into a 3-day-old pupa induced Vg in the hemolymph 4 days earlier than in the control. Methoprene also induced a faster decline in BP-A and BP-B titers in the hemolymph with a corresponding increase of the Vg titer. These results suggest that juvenile hormone (JH) induces not only vitellogenesis but also the uptake of these proteins by stimulating the metamorphosis of fat body during the pupal stage.

The Feeding Experiments End-user Database (FEED) is a research tool developed by the Mammalian Feeding Working Group at the National Evolutionary Synthesis Center that permits synthetic, evolutionary analyses of the physiology of mammalian feeding. The tasks of the Working Group are to compile physiologic data sets into a uniform digital format stored at a central source, develop a standardized terminology for describing and organizing the data, and carry out a set of novel analyses using FEED. FEED contains raw physiologic data linked to extensive metadata. It serves as an archive for a large number of existing data sets and a repository for future data sets. The metadata are stored as text and images that describe experimental protocols, research subjects, and anatomical information. The metadata incorporate controlled vocabularies to allow consistent use of the terms used to describe and organize the physiologic data. The planned analyses address long-standing questions concerning the phylogenetic distribution of phenotypes involving muscle anatomy and feeding physiology among mammals, the presence and nature of motor pattern conservation in the mammalian feeding muscles, and the extent to which suckling constrains the evolution of feeding behavior in adult mammals. We expect FEED to be a growing digital archive that will facilitate new research into understanding the evolution of feeding anatomy. PMID:21700574

Since the announcement of the ban on the use of antibiotics as antimicrobial growth promoters in the feed of pigs in 2006 the investigation towards alternative feed additives has augmented considerably. Although fermented liquid feed is not an additive, but a feeding strategy, the experimental work examining its possible advantages also saw a rise. The use of fermented liquid feed (FLF) has two main advantages, namely that the simultaneous provision of feed and water may result in an alleviation of the transition from the sow milk to solid feed and may also reduce the time spent to find both sources of nutrients, and secondly, that offering FLF with a low pH may strengthen the potential of the stomach as a first line of defence against possible pathogenic infections. Because of these two advantages, FLF is often stated as an ideal feed for weaned piglets. The results obtained so far are rather variable, but in general they show a better body weight gain and worse feed/gain ratio for the piglets. However, for growing-finishing pigs on average a better feed/gain ratio is found compared to pigs fed dry feed. This better performance is mostly associated with less harmful microbiota and better gut morphology. This review provides an overview of the current knowledge of FLF for pigs,dealing with the FLF itself as well as its effect on the gastrointestinal tract and animal performance.

In the current investigation, we evaluated a method for increasing self-feeding with 3 children with a history of food refusal. The children never (2 children) or rarely (1 child) self-fed bites of food when the choice was between self-feeding and escape from eating. When the choice was between self-feeding 1 bite of food or being fed an identical bite of food, self-feeding was low (2 children) or variable (1 child). Levels of self-feeding increased for 2 children when the choice was between self-feeding 1 bite of food or being fed multiple bites of the same food. For the 3rd child, self-feeding increased when the choice was between self-feeding 1 bite of food or being fed multiple bites of a less preferred food. The results showed that altering the contingencies associated with being fed increased the probability of self-feeding, but the specific manipulations that produced self-feeding were unique to each child.

Too often the equipment used to move difficult to handle powdery material from a hopper to process creates a complex of devices whose end result falls short on good performance...simply because equipment design, in many cases, has not kept up with advanced technological concepts in fine powder handling. The Feeder, being the key to an efficient feed system, must be assured a continuous flow from the storage tank...without bridging, arching, spasmodic flow or uncontrolled flushing. The causes and effects of flow problems are discussed and also solutions are offered based on the combination of theoretical and practical experience.

Humans have benefited from the unique catalytic properties of enzymes, in particular for food production, for thousands of years. Prominent examples include the production of fermented alcoholic beverages, such as beer and wine, as well as bakery and dairy products. The chapter reviews the historic background of the development of modern enzyme technology and provides an overview of the industrial food and feed enzymes currently available on the world market. The chapter highlights enzyme applications for the improvement of resource efficiency, the biopreservation of food, and the treatment of food intolerances. Further topics address the improvement of food safety and food quality.

The fall armyworm Spodoptera frugiperda is a prime noctuid pest of maize on the American continents where it has remained confined despite occasional interceptions by European quarantine services in recent years. The pest has currently become a new invasive species in West and Central Africa where outbreaks were recorded for the first time in early 2016. The presence of at least two distinct haplotypes within samples collected on maize in Nigeria and São Tomé suggests multiple introductions into the African continent. Implications of this new threat to the maize crop in tropical Africa are briefly discussed.

The fall armyworm Spodoptera frugiperda is a prime noctuid pest of maize on the American continents where it has remained confined despite occasional interceptions by European quarantine services in recent years. The pest has currently become a new invasive species in West and Central Africa where outbreaks were recorded for the first time in early 2016. The presence of at least two distinct haplotypes within samples collected on maize in Nigeria and São Tomé suggests multiple introductions into the African continent. Implications of this new threat to the maize crop in tropical Africa are briefly discussed. PMID:27788251

The resistance of fall armyworm (FAW), Spodoptera frugiperda, has been characterized to some Cry and Vip3A proteins of Bacillus thuringiensis (Bt) expressed in transgenic maize in Brazil. Here we evaluated the effective dominance of resistance based on the survival of neonates from selected Bt-resistant, heterozygous, and susceptible (Sus) strains of FAW on different Bt maize and cotton varieties. High survival of strains resistant to the Cry1F (HX-R), Cry1A.105/Cry2Ab (VT-R) and Cry1A.105/Cry2Ab/Cry1F (PW-R) proteins was detected on Herculex, YieldGard VT PRO and PowerCore maize. Our Vip3A-resistant strain (Vip-R) exhibited high survival on Herculex, Agrisure Viptera and Agrisure Viptera 3 maize. However, the heterozygous from HX-R × Sus, VT-R × Sus, PW-R × Sus and Vip-R × Sus had complete mortality on YieldGard VT PRO, PowerCore, Agrisure Viptera, and Agrisure Viptera 3, whereas the HX-R × Sus and Vip-R × Sus strains survived on Herculex maize. On Bt cotton, the HX-R, VT-R and PW-R strains exhibited high survival on Bollgard II. All resistant strains survived on WideStrike, but only PW-R and Vip-R × Sus survived on TwinLink. Our study provides useful data to aid in the understanding of the effectiveness of the refuge strategy for Insect Resistance Management of Bt plants.

A methanol extract from the roots and aerial parts of Myrtillocactus geometrizans (Cactaceae) yielded peniocerol 1, macdougallin 2, and chichipegenin 3. The natural products 1, 2 their mixtures, MeOH and CH(2)Cl(2) extracts showed insecticidal and insect growth regulatory activity against fall armyworm [Spodoptera frugiperda J. E. Smith (Lepidoptera: Noctuidae)], an important insect pest of corn, and [Tenebrio molitor (Coleoptera)], a pest of stored grains in Mexico. The most active compounds were 1, 2, and a mixture (M(2)) of 1 and 2 (6:4). All these extracts, compounds and the mixture had insect growth regulating (IGR) activity between 5.0 and 50.0 ppm and insecticidal effects between 50 and 300 ppm in diets. The extracts were insecticidal to larvae, with lethal doses between 100 and 200 ppm. These compounds appear to have selective effects on the pre-emergence metabolism of Coleoptera, because in all treatments of the larvae of T. molitor, pupation were shortened and this process show precociousness in relation to controls. In contrast to S. frugiperda larvae, onset of pupation was noticeably delayed. Emergence in both cases was drastically diminished. In both pupae and in the few adults that were able to emerge, many deformations were observed. The results of these assays indicated that the compounds were more active than other known natural insect growth inhibitors such as gedunin and methanol extracts of Cedrela salvadorensis and Yucca periculosa. Peniocerol, macdougallin and chichipegenin, as well as mixtures of these substances, may be useful as natural insecticidal agents.

The current identification of microRNAs (miRNAs) in insects is largely dependent on genome sequences. However, the lack of available genome sequences inhibits the identification of miRNAs in various insect species. In this study, we used a miRNA database of the silkworm Bombyx mori as a reference to identify miRNAs in Helicoverpa armigera and Spodoptera litura using deep sequencing and homology analysis. Because all three species belong to the Lepidoptera, the experiment produced reliable results. Our study identified 97 and 91 conserved miRNAs in H. armigera and S. litura, respectively. Using the genome of B. mori and BAC sequences of H. armigera as references, 1 novel miRNA and 8 novel miRNA candidates were identified in H. armigera, and 4 novel miRNA candidates were identified in S. litura. An evolutionary analysis revealed that most of the identified miRNAs were insect-specific, and more than 20 miRNAs were Lepidoptera-specific. The investigation of the expression patterns of miR-2a, miR-34, miR-2796-3p and miR-11 revealed their potential roles in insect development. miRNA target prediction revealed that conserved miRNA target sites exist in various genes in the 3 species. Conserved miRNA target sites for the Hsp90 gene among the 3 species were validated in the mammalian 293T cell line using a dual-luciferase reporter assay. Our study provides a new approach with which to identify miRNAs in insects lacking genome information and contributes to the functional analysis of insect miRNAs. PMID:23289012

Camptothecin (CPT), a plant alkaloid originally isolated from the native Chinese tree, Camptotheca acuminate, exerts the toxic effect by targeting eukaryotic DNA topoisomerase 1 (DNA Topo1). Besides as potent anti-cancer agents, CPT and its derivatives are now being explored as potential pesticides for insect control. In this study, we assessed their toxicity to an insect homolog, the Topo1 protein from beet armyworms (Spodoptera exigua Hübner), a worldwide pest of many important crops. The S. exigua Topo1 gene contains an ORF of 2790 base pairs that is predicted to encode a polypeptide of 930 amino acids. The deduced polypeptide exhibits polymorphism at residue sites V420, L530, A653 and T729 (numbered according to human Topo1) among insect species, which are predicted to confer sensitivity to CPT. The DNA relaxation activity of this protein was subsequently examined using a truncated form that contained the residues 337–930 and was expressed in bacteria BL21 cells. The purified protein retained the ability to relax double-stranded DNA and was susceptible to CPT and its derivative hydroxy-camptothecin (HCPT) in a dose-dependent manner. The same inhibitory effect was also found on the native Topo1 extracted from IOZCAS-Spex-II cells, a cell line established from beet armyworms. Additionally, CPT and HCPT treatment reduced the steady accumulation of Topo1 protein despite the increased mRNA expression in response to the treatment. Our studies provide information of the S. exigua Topo1 gene and its amino acid polymorphism in insects and uncover some clues about potential mechanisms of CPT toxicity against insect pests. These results also are useful for development of more effective Topo1-targeted CPT insecticides in the future. PMID:23451051

Antimicrobial peptides (AMPs) and lysozymes are the main effectors of the insect immune system, and they are involved in both local and systemic responses. Among local responses, midgut immune reaction plays an important role in fighting pathogens that reach the insect body through the oral route, as do many microorganisms used in pest control. Under this point of view, understanding how insects defend themselves locally during the first phases of infections caused by food-borne pathogens is important to further improve microbial control strategies. In the present study, we analyzed the transcriptional response of AMPs and lysozymes in the midgut of Spodoptera exigua (Lepidoptera: Noctuidae), a polyphagous pest that is commonly controlled by products based on Bacillus thuringiensis (Bt) or baculovirus. First, we comprehensively characterized the transcripts encoding AMPs and lysozymes expressed in S. exigua larval midgut, identifying 35 transcripts that represent the S. exigua arsenal against microbial infection. Secondly, we analyzed their expression in the midgut after ingestion of sub-lethal doses of two different pore-forming B. thuringiensis toxins, Cry1Ca and Vip3Aa, and the S. exigua nucleopolyhedrovirus (SeMNPV). We observed that both Bt toxins triggered a similar, wide and in some cases high transcriptional activation of genes encoding AMPs and lysozymes, which was not reflected in the activation of the classical systemic immune-marker phenoloxidase in hemolymph. Baculovirus ingestion resulted in the opposed reaction: Almost all transcripts coding for AMPs and lysozymes were down-regulated or not induced 96 hours post infection. Our results shed light on midgut response to different virulence factors or pathogens used nowadays as microbial control agents and point out the importance of the midgut immune response contribution to the larval immunity.

Baculoviruses are large DNA viruses that infect insect species such as Lepidoptera and are used in biotechnology for protein production and in agriculture as insecticides against crop pests. Baculoviruses require activity of host proteasomes for efficient reproduction, but how they control the cellular proteome and interact with the ubiquitin proteasome system (UPS) of infected cells remains unknown. In this report, we analyzed possible changes in the subunit composition of 26S proteasomes of the fall armyworm, Spodoptera frugiperda (Sf9), cells in the course of infection with the Autographa californica multiple nucleopolyhedrovirus (AcMNPV). 26S proteasomes were purified from Sf9 cells by an immune affinity method and subjected to 2D gel electrophoresis followed by MALDI-TOF mass spectrometry and Mascot search in bioinformatics databases. A total of 34 homologues of 26S proteasome subunits of eukaryotic species were identified including 14 subunits of the 20S core particle (7 α and 7 β subunits) and 20 subunits of the 19S regulatory particle (RP). The RP contained homologues of 11 of RPN-type and 6 of RPT-type subunits, 2 deubiquitinating enzymes (UCH-14/UBP6 and UCH-L5/UCH37), and thioredoxin. Similar 2D-gel maps of 26S proteasomes purified from uninfected and AcMNPV-infected cells at 48hpi confirmed the structural integrity of the 26S proteasome in insect cells during baculovirus infection. However, subtle changes in minor forms of some proteasome subunits were detected. A portion of the α5(zeta) cellular pool that presumably was not associated with the proteasome underwent partial proteolysis at a late stage in infection.

To gain insight into the comparative function in stress response of HSPs in insects, three HSP cDNAs were cloned from the fat body of the beet armyworm Spodoptera exigua (Lepidoptera, Noctuidae). SexHSP70, SexHSP74 and SexHSP83 cDNAs encoding the protein of 667, 685 and 717 amino acids, with the pI of 5.52, 5.75 and 5.02, respectively. Northern blotting revealed that all three SexHSP mRNAs are expressed in the fat body, mid-gut, spermary and tracheae. SexHSP70, SexHSP74and SexHSP83 mRNAs were expressed in the fat body and whole body at different levels during different developmental stages. The three SexHSP transcripts were highly expressed in the fat body on the first day of fifth instar larvae, on the fourth and seventh days of the pupa stage, and in the whole body on the initial stages of larvae. Under heat and cold shock conditions, SexHSP70 and SexHSP83 mainly functioned during heat shock and cooling and SexHSP83 also had a function in the recovery stage. SexHSP74 had important functions in short-term heat shock and recovery, as well as long-term cooling. The results revealed that long-term shocking can affect SexHSP74 and SexHSP83 expression and long-term cooling can influence SexHSP83 expression during the recovery stage.

Insects can increase their resistance to cold stress by prior exposure to non-lethal cold temperatures. Here, we investigated the supercooling capacity and survival of eggs, 3rd and 5th instar larvae, and pupae of Spodoptera exigua (Lepidoptera: Noctuidae) during CA, and responses to various pre-treatment protocols, including constant temperatures, thermoperiods, and RCH, RHH, RCH+RHH and RHH+RCH combined with thermoperiods. Only acclimated eggs demonstrated a significant decrease in SCP, from -20.7±0.3 to -22.9±0.3°C, among all experimental groups compared to non-acclimated stages. Survival increased by 17.5% for eggs, 40.0% and 13.3% for 3rd and 5th instar larvae, and by 20.0% for pupae after CA. Compared to controls, survival of eggs under the conditions of thermoperiod (5:15°C), thermoperiod (5:15°C)+RHH, and thermoperiod (5:15, 10:20, and 15:25°C)+RCH significantly increased. In addition, survival of 3rd and 5th instar larvae and pupae increased under the conditions of thermoperiod (5:15°C) and thermoperiod (5:15°C)+RCH, possibly due to the induction of heat shock proteins or cryoprotectants. However, the pre-treatments of thermoperiod+RCH+RHH and thermoperiod+RHH+RCH did not significantly enhance survival of any developmental stage. These adaptive responses may allow S. exigua to enhance supercooling capacity and survival in response to seasonal or unexpected diurnal decreases in environmental temperatures.

Antimicrobial peptides (AMPs) and lysozymes are the main effectors of the insect immune system, and they are involved in both local and systemic responses. Among local responses, midgut immune reaction plays an important role in fighting pathogens that reach the insect body through the oral route, as do many microorganisms used in pest control. Under this point of view, understanding how insects defend themselves locally during the first phases of infections caused by food-borne pathogens is important to further improve microbial control strategies. In the present study, we analyzed the transcriptional response of AMPs and lysozymes in the midgut of Spodoptera exigua (Lepidoptera: Noctuidae), a polyphagous pest that is commonly controlled by products based on Bacillus thuringiensis (Bt) or baculovirus. First, we comprehensively characterized the transcripts encoding AMPs and lysozymes expressed in S. exigua larval midgut, identifying 35 transcripts that represent the S. exigua arsenal against microbial infection. Secondly, we analyzed their expression in the midgut after ingestion of sub-lethal doses of two different pore-forming B. thuringiensis toxins, Cry1Ca and Vip3Aa, and the S. exigua nucleopolyhedrovirus (SeMNPV). We observed that both Bt toxins triggered a similar, wide and in some cases high transcriptional activation of genes encoding AMPs and lysozymes, which was not reflected in the activation of the classical systemic immune-marker phenoloxidase in hemolymph. Baculovirus ingestion resulted in the opposed reaction: Almost all transcripts coding for AMPs and lysozymes were down-regulated or not induced 96 hours post infection. Our results shed light on midgut response to different virulence factors or pathogens used nowadays as microbial control agents and point out the importance of the midgut immune response contribution to the larval immunity. PMID:25993013

To enhance the toxicity of the Bacillus thuringiensis subsp. aizawai strain SP41 (SP41), the vegetative insecticidal protein (Vip) gene vip3A from SP41 was redirected to the sporulation stage by replacing its native promoter with the strong promoter P19 of the cry11Aa operon. Compared to the wild type, SP41 with PVIP (vip3A with its native promoter and ter) had the relative expression ratios of 457, 548, and 290 at 8, 14, and 20 h of cultivation, respectively, as measured by quantitative reverse transcription polymerase chain reaction (PCR). SP41 transformed by P19VIP (vip3A controlled by P19 promoter with vip3A ter) showed higher expressions (23, 2055, 1831) at the same time points. SP41 with P19VIP20 (vip3A controlled by P19 promoter and containing P20 and operon ter) had the lowest expression levels (3, 11, 9) at any time point. SDS-PAGE analysis of proteins in the culture supernatant of the P19VIP at 8, 14, and 20 h demonstrated a significant increase in Vip3A at the sporulation stage. Using the surface contamination bioassay, the 50% lethal concentration (LC(50)) of whole culture of PVIP, P19VIP, and P19VIP20 at 20 and 48 h of cultivation against Spodoptera exigua larvae were (68.3, 21.2, and 60.2 microg cm(-2)) and (69.8, 41.8, and 74.6 microg cm(-2)), respectively, compared with 86.6 and 104.4 microg cm(-2) for SP41. The results showed that Vip from P19VIP, expressed at spore stage at 20 and 48 h, can increase the toxicity of SP41 for 4.1- and 2.5-fold, respectively.

The resistance of fall armyworm (FAW), Spodoptera frugiperda, has been characterized to some Cry and Vip3A proteins of Bacillus thuringiensis (Bt) expressed in transgenic maize in Brazil. Here we evaluated the effective dominance of resistance based on the survival of neonates from selected Bt-resistant, heterozygous, and susceptible (Sus) strains of FAW on different Bt maize and cotton varieties. High survival of strains resistant to the Cry1F (HX-R), Cry1A.105/Cry2Ab (VT-R) and Cry1A.105/Cry2Ab/Cry1F (PW-R) proteins was detected on Herculex, YieldGard VT PRO and PowerCore maize. Our Vip3A-resistant strain (Vip-R) exhibited high survival on Herculex, Agrisure Viptera and Agrisure Viptera 3 maize. However, the heterozygous from HX-R × Sus, VT-R × Sus, PW-R × Sus and Vip-R × Sus had complete mortality on YieldGard VT PRO, PowerCore, Agrisure Viptera, and Agrisure Viptera 3, whereas the HX-R × Sus and Vip-R × Sus strains survived on Herculex maize. On Bt cotton, the HX-R, VT-R and PW-R strains exhibited high survival on Bollgard II. All resistant strains survived on WideStrike, but only PW-R and Vip-R × Sus survived on TwinLink. Our study provides useful data to aid in the understanding of the effectiveness of the refuge strategy for Insect Resistance Management of Bt plants.

Transgenic maize, Zea maize L., expressing the Cry1F protein from Bacillus thuringiensis has been registered for Spodoptera frugiperda (J. E. Smith) control since 2003. Unexpected damage to Cry1F maize was reported in 2006 in Puerto Rico and Cry1F resistance in S. frugiperda was documented. The inheritance of Cry1F resistance was characterized in a S. frugiperda resistant strain originating from Puerto Rico, which displayed >289-fold resistance to purified Cry1F. Concentration-response bioassays of reciprocal crosses of resistant and susceptible parental populations indicated that resistance is recessive and autosomal. Bioassays of the backcross of the F1 generation crossed with the resistant parental strain suggest that a single locus is responsible for resistance. In addition, cross-resistance to Cry1Aa, Cry1Ab, Cry1Ac, Cry1Ba, Cry2Aa and Vip3Aa was assessed in the Cry1F-resistant strain. There was no significant cross-resistance to Cry1Aa, Cry1Ba and Cry2Aa, although only limited effects were observed in the susceptible strain. Vip3Aa was highly effective against susceptible and resistant insects indicating no cross-resistance with Cry1F. In contrast, low levels of cross-resistance were observed for both Cry1Ab and Cry1Ac. Because the resistance is recessive and conferred by a single locus, an F1 screening assay was used to measure the frequency of Cry1F-resistant alleles from populations of Florida and Texas in 2010 and 2011. A total frequency of resistant alleles of 0.13 and 0.02 was found for Florida and Texas populations, respectively, indicating resistant alleles could be found in US populations, although there have been no reports of reduced efficacy of Cry1F-expressing plants.

The resistance of fall armyworm (FAW), Spodoptera frugiperda, has been characterized to some Cry and Vip3A proteins of Bacillus thuringiensis (Bt) expressed in transgenic maize in Brazil. Here we evaluated the effective dominance of resistance based on the survival of neonates from selected Bt-resistant, heterozygous, and susceptible (Sus) strains of FAW on different Bt maize and cotton varieties. High survival of strains resistant to the Cry1F (HX-R), Cry1A.105/Cry2Ab (VT-R) and Cry1A.105/Cry2Ab/Cry1F (PW-R) proteins was detected on Herculex, YieldGard VT PRO and PowerCore maize. Our Vip3A-resistant strain (Vip-R) exhibited high survival on Herculex, Agrisure Viptera and Agrisure Viptera 3 maize. However, the heterozygous from HX-R × Sus, VT-R × Sus, PW-R × Sus and Vip-R × Sus had complete mortality on YieldGard VT PRO, PowerCore, Agrisure Viptera, and Agrisure Viptera 3, whereas the HX-R × Sus and Vip-R × Sus strains survived on Herculex maize. On Bt cotton, the HX-R, VT-R and PW-R strains exhibited high survival on Bollgard II. All resistant strains survived on WideStrike, but only PW-R and Vip-R × Sus survived on TwinLink. Our study provides useful data to aid in the understanding of the effectiveness of the refuge strategy for Insect Resistance Management of Bt plants. PMID:27721425

When cutworm Spodoptera litura larvae were fed on the diets with different lead (Pb) concentrations for one or five generations, changes in growth and food utilization were recorded; Pb accumulations were detected by Atomic Absorption Spectrophotometer; changes in midgut cell ultrastructure were observed by Transmission Electron Microscopy (TEM). The effects of Pb stress on S. litura growth and food utilization differed significantly between insects of the 1st and 5th generation. The male-female rate of 200mgkg(-1) Pb treatment from the 1st generation and 50mgkg(-1) Pb treatment from the 5th generation was significantly higher than control. No significant difference of Pb accumulations was found in larvae, pupae and adults between the 1st and 5th generation. No significant difference of Pb accumulations in corresponding tissues of larvae was found between male and female. Compared to fat body, hemolymph, head, foregut and hindgut, the highest Pb accumulation was found in migut of larvae exposed to 200mgkg(-1) Pb. TEM showed that expanded intercellular spaces were observed in Pb-treated midgut cells. The nuclei were strongly destroyed by Pb stress, evidenced by chromatin condensation and destroyed nuclear envelope. Mitochondria became swollen with some broken cristae after exposure to Pb. Therefore, neither gender nor progeny difference was present in Pb accumulations of S. litura, although effects of Pb stress on S. litura growth and food utilization differed from different generations and genders. Pb accumulations in midgut caused pathological changes in cells ultrastructure, possibly reflected the growth and food utilization of S. litura.

ABSTRACT Cleft lip and palate patients have all rights like other normal individuals, to enjoy the benefits of nourishment. Knowledge has to be there about the different feeding positions like straddle, dancer hand position along with the use of specially designed bottles and nipples. Parent's should be trained about the correct positions of feeding, in extreme of the cases in which parents are not able to follow these instructions, feeding obturators can be given. How to cite this article: Jindal MK, Khan SY. How to Feed Cleft Patient? Int J Clin Pediatr Dent 2013;6(2):100-103. PMID:25206201

A device for providing uniform powder flow to the nozzles when creating solid structures using a solid fabrication system such as the directed light fabrication (DLF) process. In the DLF process, gas entrained powders are passed through the focal point of a moving high-power laser light which fuses the particles in the powder to a surface being built up in layers. The invention is a device providing uniform flow of gas entrained powders to the nozzles of the DLF system. The device comprises a series of modular splitters which are slidably interconnected and contain an integral flow control mechanism. The device can take the gas entrained powder from between one to four hoppers and split the flow into eight tubular lines which feed the powder delivery nozzles of the DLF system.

A device for providing uniform powder flow to the nozzles when creating solid structures using a solid fabrication system such as the directed light fabrication (DLF) process. In the DLF process, gas entrained powders are passed through the focal point of a moving high-power laser light which fuses the particles in the powder to a surface being built up in layers. The invention is a device providing uniform flow of gas entrained powders to the nozzles of the DLF system. The device comprises a series of modular splitters which are slidably interconnected and contain an integral flow control mechanism. The device can take the gas entrained powder from between one to four hoppers and split the flow into eight tubular lines which feed the powder delivery nozzles of the DLF system.

Robust sleep/wake rhythms are important for health and cognitive function. Unfortunately, many people are living in an environment where their circadian system is challenged by inappropriate meal- or work-times. Here we scheduled food access to the sleep time and examined the impact on learning and memory in mice. Under these conditions, we demonstrate that the molecular clock in the master pacemaker, the suprachiasmatic nucleus (SCN), is unaltered while the molecular clock in the hippocampus is synchronized by the timing of food availability. This chronic circadian misalignment causes reduced hippocampal long term potentiation and total CREB expression. Importantly this mis-timed feeding resulted in dramatic deficits in hippocampal-dependent learning and memory. Our findings suggest that the timing of meals have far-reaching effects on hippocampal physiology and learned behaviour. DOI: http://dx.doi.org/10.7554/eLife.09460.001 PMID:26652002

This article provides an overview of future prospects for feeding the world's growing population. The discussion focuses on obstacles such as limited agricultural land, degraded soil and water, and water shortages. The evidence suggests that sustainability is declining, especially in poor, food-deficit countries with growing populations. The world is segregated into the haves, the poor have-nots, and the rich have-nots. North America, Europe, and Australia have enough cropland to feed their populations. The poor have-nots are located mostly in sub-Saharan Africa, 7 countries each in the Middle East and Latin America, 6 in Oceania, and the rest in Central and South Asia. The poor have-nots amount to 3 billion out of 6 billion total population. The rich have-nots include countries such as Japan and Singapore, plus China, Indonesia, Peru, Chile, and Saudi Arabia and other Gulf states. The rich have-nots must import food. The world grain harvest is no longer tripling. Per person yields have declined. Increasing food productivity must rely on existing lands. The size of family farms has declined. Almost 2 billion hectares of crop and grazing land is degraded. Yields from irrigated land that are 33% of world food supply have declined. In 1990, 28 countries with 335 million people faced chronic water shortages or scarcity. Water is being polluted. Fish stocks are being depleted. Genetic diversity is being lost. In 182 food deficit countries, population growth must be slowed, and agriculture must be sustainable. Food is neither produced nor consumed equitably. Malnutrition is caused by poverty. Food security cannot be achieved if land and water become increasingly degraded or lost.

Populations of Eruca sativa (Brassicaceae) from desert and Mediterranean (Med) habitats in Israel differ in their defense against larvae of the generalist Spodopteralittoralis but not the specialist Pieris brassicae. Larvae of the generalist insect feeding on plants of the Med population gained significantly less weight than those feeding on the desert plants, and exogenous application of methyl jasmonate (MJ) on leaves of the Med plants significantly reduced the level of damage created by the generalist larvae. However, MJ treatment significantly induced resistance in plants of the desert population, whereas the generalist larvae caused similar damage to MJ-induced and noninduced plants. Analyses of glucosinolates and expression of genes in their synthesis pathway indicated that defense in plants of the Med population against the generalist insect is governed by the accumulation of glucosinolates. In plants of the desert population, trypsin proteinase inhibitor activity was highly induced in response to herbivory by S. littoralis. Analysis of genes in the defense-regulating signaling pathways suggested that in response to herbivory, differences between populations in the induced levels of jasmonic acid, ethylene, and salicylic acid mediate the differential defenses against the insect. In addition, expression analysis of myrosinase-associated protein NSP2 suggested that in plants of the desert population, glucosinolates breakdown products were primarily directed to nitrile production. We suggest that proteinase inhibitors provide an effective defense in the desert plants, in which glucosinolate production is directed to the less toxic nitriles. The ecological role of nitrile production in preventing infestation by specialists is discussed.

We evaluated the anti-insectan activity of extracts from different vegetative parts of ten plant species native to Uruguay. The selected plants belong to five families: Bignoniaceae: Clytostoma callistegioides, Dolichandra cynanchoides, Macfadyena unguis-cati; Sapindaceae: Dodonaea viscosa, Allophylus edulis, Serjania meridionalis; Lamiaceae: Salvia procurrens, Salvia guaranitica; Solanaceae: Lycium cestroides; and Phytolaccaceae: Phytolacca dioica. The extracts were evaluated in independent bioassays against four insect pests and one beneficial insect. Aphid settling inhibition was evaluated with a grass specialist, Rhopalosiphum padi, and a feeding generalist, Myzus persicae (both Hemiptera: Aphididae). Antifeedant activity was tested with adults of the specialist Epilachna paenulata (Coleoptera: Coccinellidae) and larvae of the generalist Spodopteralittoralis (Lepidoptera: Noctuidae). Finally, contact toxicity was assessed with honey bees, Apis mellifera (Hymenoptera: Apidae). Strong settling inhibition (SI) activity (expressed as %SI, where 100% means complete inhibition by the extract) was found only for the twig extracts of A. edulis (Sapindaceae) against M. persicae (% SI = 77 +/- 4). Antifeedant activity (expressed as % of feeding reduction (FR), where 100% means no consumption on extract-treated diet) against E. paenulata was significant for the leaf extracts of L. cestroides (Solanaceae) (% FR = 100 +/- 0) as well as of all Bignoniaceae and Sapindaceae species. No extracts were active against S. littoralis larvae, and most of them were innocuous to honey bees, with the exception of L. cestroides and S. meridionalis leaf extracts.

The cell lytic activity and toxicity against lepidopteran larvae of 13 venom peptides (4 OdVPs and 9 EpVPs) from two solitary hunting wasps, Orancistrocerus drewseni and Eumenes pomiformis, were examined with mastoparan as a reference peptide. Of the 13 peptides, 7 were predicted to have α-helical structures that exhibit the typical character of amphipathic α-helical antimicrobial peptides. The remaining peptides exhibited coil structures; among these, EpVP5 possesses two Cys residues that form an internal disulfide bridge. All the helical peptides including mastoparan showed antimicrobial and insect cell lytic activities, whereas only two of them were hemolytic against human erythrocytes. The helical peptides induced a feeding disorder when injected into the vicinity of the head and thorax of Spodoptera exigua larvae, perhaps because their non-specific neurotoxic or myotoxic action induced cell lysis. At low concentrations, however, these helical peptides increased cell permeability without inducing cell lysis. These findings suggest that the helical venom peptides may function as non-specific neurotoxins or myotoxins and venom-spreading factors at low concentrations, as well as preservatives for long-term storage of the prey via antimicrobial, particularly antifungal, activities.

The fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), is a target of transgenic maize and cotton expressing Bacillus thuringiensis (Bt) proteins in both North and South Americas. In the falls of 2013 and 2014, a total of 215 F2 two-parent families of S. frugiperda were es...

BACKGROUND: Insect cytochrome P450 monooxygenases (CYPs or P450s) play an important role in detoxifying insecticides leading to resistance in insect populations. A polyphagous pest, Spodoptera litura (Lepidoptera, Noctuidae) has been shown to be resistant to a wide range of insecticides. In this stu...

Herbivore-induced plant volatiles (HIPVs) serve as specific cues to higher trophic levels. Novel, exotic herbivores entering native foodwebs may disrupt the infochemical network as a result of changes in HIPV profiles. Here, we analysed HIPV blends of native Brassica rapa plants infested with one of 10 herbivore species with different coexistence histories, diet breadths and feeding modes. Partial least squares (PLS) models were fitted to assess whether HIPV blends emitted by Dutch B. rapa differ between native and exotic herbivores, between specialists and generalists, and between piercing-sucking and chewing herbivores. These models were used to predict the status of two additional herbivores. We found that HIPV blends predicted the evolutionary history, diet breadth and feeding mode of the herbivore with an accuracy of 80% or higher. Based on the HIPVs, the PLS models reliably predicted that Trichoplusia ni and Spodoptera exigua are perceived as exotic, leaf-chewing generalists by Dutch B. rapa plants. These results indicate that there are consistent and predictable differences in HIPV blends depending on global herbivore characteristics, including coexistence history. Consequently, native organisms may be able to rapidly adapt to potentially disruptive effects of exotic herbivores on the infochemical network.

A leucine aminopeptidase (EC 3.4.11.1) cDNA clone (DR57) that was induced in response to Pseudomonas syringae pv. tomato (P.s. tomato) infection was isolated using a subtractive hybridization-enriched cDNA probe. Genomic DNA blot analysis showed that the tomato genome had two leucine aminopeptidase genes. The levels of DR57 mRNAs after P.s. tomato infection and mechanical wounding were determined in two inbred tomato lines that exhibit susceptibility and resistance to P.s. tomato. DR57 mRNAs were detected 12 hours after infection and 4 hours after wounding. Furthermore, DR57 mRNAs were systemically induced in response to wounding. DR57 mRNAs were induced in leaves after Spodopteralittoralisfeeding but were not detected in detached leaf controls. Possible roles for the DR57 leucine aminopeptidase in the defense reactions are discussed.

development, assembly and test of an electric propulsion xenon feed system for a flight technology demonstration program. Major accomplishments...pressure transducer feedback, the PFCV has successfully fed xenon to a 200 watt Hall Effect Thruster in a Technology Demonstration Program. The feed

The ultimate objective of an aquaculture feed manufacturer and aquaculture food supplier is to ensure that the feed or food produced is both safe and wholesome. Reported food safety risks, which may be associated with the use of commercial animal feeds, including compound aquaculture feeds, usually result from the possible presence of unwanted contaminants, either within the feed ingredients used or from the external contamination of the finished feed on prolonged storage. The major animal feed contaminants that have been reported to date have included Salmonellae, mycotoxins, veterinary drug residues, persistent organic pollutants, agricultural and other chemicals (solvent residues, melamine), heavy metals (mercury, lead, cadmium) and excess mineral salts (hexavalent chromium, arsenic, selenium, flourine), and transmissible spongiform encephalopathies. Apart from the direct negative effect of these possible contaminants on the health of the cultured target species, there is a risk that the feed contaminants may be passed along the food chain, via contaminated aquaculture produce, to consumers. In recent years, public concern regarding food safety has increased as a consequence of the increasing prevalence of antibiotic residues, persistent organic pollutants, and chemicals in farmed seafood. The important role played by the Food and Agriculture Organization of the United Nations (FAO) and the Codex Alimentarius Commission in the development of international standards, guidelines, and recommendations to protect the health of consumers and ensure fair practices in the food trade is discussed.

Compositional analysis is central to determining the nutritional value of feedstuffs. The utility of the values and how they should be used depends on how representative the feed subsample is, the nutritional relevance of the assays, analytical variability of the analyses, and whether a feed is suit...

To determine the effects of enteral tube feeding on the incidence of pneumonia, we performed a retrospective review of all clients at our institution who had gastrostomy or jejunostomy tubes placed over a 10-year period. Ninety-three subjects had a history of pneumonia before feeding tube insertion. Eighty had gastrostomy and 13, jejunostomy…

The historical evolution of infant feeding includes wet nursing, the feeding bottle, and formula use. Before the invention of bottles and formula, wet nursing was the safest and most common alternative to the natural mother's breastmilk. Society's negative view of wet nursing, combined with improvements of the feeding bottle, the availability of animal's milk, and advances in formula development, gradually led to the substitution of artificial feeding for wet nursing. In addition, the advertising and safety of formula products increased their popularity and use among society. Currently, infant formula-feeding is widely practiced in the United States and appears to contribute to the development of several common childhood illnesses, including atopy, diabetes mellitus, and childhood obesity. PMID:20190854

Transplantation affords recipients the potential for a full life and, for some, parenthood. Female transplant recipients must continue to take immunosuppression during pregnancy and breast-feeding. This article reviews case and series reports regarding breast-feeding in those taking transplant medications. Avoidance of breast-feeding has been the customary advice because of the potential adverse effects of immunosuppressive exposure on the infant. Subsequent studies have demonstrated that not all medication exposure translates to risk for the infant, that the exposure in utero is greater than via breast milk and that no lingering effects due to breast-feeding have been found to date in infants who were breast-fed while their mothers were taking prednisone, azathioprine, cyclosporine, and/or tacrolimus. Thus, except for those medications where clinical information is inadequate (mycophenolic acid products, sirolimus, everolimus, and belatacept), the recommendation for transplant recipients regarding breast-feeding has evolved into one that is cautiously optimistic.

The evidence linking bottle feeding to infant and early childhood mortality has been reviewed. Ecological studies of national time trends in infant mortality do not parallel breast feeding trends in those countries, and indicate that falling death rates are more likely to be related to better health care facilities and social conditions. Direct studies of deaths provide some contradictory findings; meta-analyses are not informative because of the many differences in statistical and sample methodology. The methodology exhibited in most studies is more likely to have over- rather than under-estimated a relationship between bottle feeding and infant mortality. Retrospective analyses must take account of changes in feeding pattern due to early signs of illness. Prospective population studies able to account for large numbers of potential confounders provide the best estimates, especially if proportional hazards models are used. Two such studies have been carried out--both showed protective effects of breast feeding.

A novel feed structure, for an antenna having a resonant electric field structure, comprising a patch element, an integrated circuit attached to the patch element, at least one inner conductor electrically connected to and terminating at the integrated circuit on a first end of the at least one inner conductor, wherein the at least one inner conductor extends through and is not electrically connected to the patch element, and wherein the at least one inner conductor is available for electrical connectivity on a second end of the at least one inner conductor, and an outer conductor electrically connected to and terminating at the patch element on a first end of the outer conductor, wherein the outer conductor is available for electrical connectivity on a second end of the outer conductor, and wherein the outer conductor concentrically surrounds the at least one inner conductor from the second end of the at least one inner conductor available for electrical connectivity to the first end of the outer conductor terminating at the patch element.

A class of feed antennas and feed antenna arrays used in the focal plane of paraboloid reflectors and exhibiting higher than normal levels of cross-polarized radiation in the diagonal planes is addressed. A model which allows prediction of element gain and aperture efficiency of the feed/reflector system is presented. The predictions are in good agreement with experimental results. Tapered slot antenna (TSA) elements are used an example of an element of this type. It is shown that TSA arrays used in multibeam systems with small beam spacings are competitive in terms of aperture efficiency with other, more standard types of arrays incorporating waveguide type elements.

Being delivered as sprays or expressed in plant, Bacillus thuringiensis (Bt) crystalline proteins (Cry toxins) display insecticidal activities against numerous Lepidopteran, Dipteran, and Coleopteran larvae. Comparative study of toxicities of Bt Cry toxins between larvae and adults may afford important new insights into the interactions of the toxins with receptor proteins in host insect, and represent intriguing targets for the control of insect pests. However, the effectiveness of Bt Cry toxins in insect adults has paid less attention. In the present article, the effectiveness of Cry1Ac and Cry1Ca on lifespans and reproductive performance of Helicoverpa armigera (Hübner) and Spodoptera exigua (Hübner) adults were evaluated by in vivo experiments. Considering transgenic plants express modified, truncated versions of cry genes yielding active toxin fragment, we used activated Bt toxins at the concentration of 500, 100, and 20 microg/ml in a 10% sucrose aquous solution. At the highest concentration, Cry1Ac and Cry1Ca shortened 48.1 and 48.9% of H. armigera female lifespan, and 43.5 and 38.5% of S. exigua female lifespan, and they reduced 37.8 and 40.3%, and 50.5 and 47.4% of H. armigera and S. exigua male lifespans respectively. Bt toxins negatively affected copulation. Exposure to 500 microg/ml of Cry1Ac and Cry1Ca greatly reduced 50.0 and 46.8%, and 58.7 and 57.3% spermatophore acceptance by H. armigera and S. exigua females, respectively. Similarly, Cry1Ac and Cry1Ca exposure decreased 40.0 and 50.3%, and 61.3 and 60.0% of spermatophore transfer by H. armigera and S. exigua males, respectively. Moreover, exposure females rather than males to 500 microg/ml of Cry1Ac and Cry1Ca significantly dropped 57.5 and 57.5% of the number of eggs laid by H. armigera, and 35.4 and 45.8% of the number of egg masses deposited by S. exigua. In contrast, both Cry1Ac and Cry1Ca did not negatively influence the egg hatchability. At the middle and the lowest concentrations, however

Spodoptera frugiperda (Sf9) ovarian cells, natural hosts for baculovirus, are good model systems to study apoptosis and also heterologous gene expression. We report that uninfected Sf9 cells readily undergo apoptosis and show increased phosphorylation of the alpha subunit of eukaryotic initiation factor 2 (eIF2alpha) in the presence of agents such as UVB light, etoposide, high concentrations of cycloheximide, and EGTA. In contrast, tunicamycin, A23187, and low concentrations of cycloheximide promoted eIF2alpha phosphorylation in Sf9 cells but without apoptosis. These findings therefore suggest that increased eIF2alpha phosphorylation does not always necessarily lead to apoptosis, but it is a characteristic hallmark of stressed cells and also of cells undergoing apoptosis. Cell death induced by the above agents was abrogated by infection of Sf9 cells with wild-type (wt) AcNPV. In contrast, Sf9 cells when infected with vAcdelta35, a virus carrying deletion of the antiapoptotic p35 gene, showed increased apoptosis and enhanced eIF2alpha phosphorylation. Further, a recombinant wt virus vAcS51D expressing human S51D, a phosphomimetic form of eIF2alpha, induced apoptosis in UVB pretreated Sf9 cells. However, infection with vAcS51A expressing a nonphosphorylatable form (S51A) of human eIF2alpha partially reduced apoptosis. Consistent with these findings, it has been observed here that caspase activation has led to increased eIF2alpha phosphorylation, while caspase inhibition by z-VAD-fmk reduced eIF2alpha phosphorylation selectively in cells exposed to proapoptotic agents. These findings therefore suggest that the stress signaling pathway determines apoptosis, and caspase activation is a prerequisite for increased eIF2alpha phosphorylation in Sf9 cells undergoing apoptosis. The findings also reinforce the conclusion for the first time that the "pancaspase inhibitor" baculovirus p35 mitigates eIF2alpha phosphorylation.

This article discusses the unequal distribution of food within and among countries of the world, poverty as the main cause of hunger, the patterns of population growth, and future prospects. The evidence reveals the potential for reaching limits of agricultural expansion. Widespread environmental destruction suggests that agricultural lands are declining. Current production patterns may not be sustainable. "Population growth is the single largest determinant of future needs." Farmers of the world's poorest regions must produce food to meet the needs of a doubled population by 2050. The 1996 World Food Summit urged that agricultural policies emphasize environmentally sustainable production methods and a wider variety of crops that would include sorghum and millet. In the early 1990s, 40% of Africa's population was food-energy deficient. Africa has some of the highest population growth rates. Fertility stands at about 6 children/woman. Even with reduced fertility, the African population is expected to double to 22% of world population by 2050. The East Asian population, which is dominated by China, is expected to reach 2.2 billion by 2050. South Asia includes some of the poorest and most densely populated countries; its population is expected to reach 2.2-3.3 billion by 2050. 43% of the malnourished population during 1990-92 lived in sub-Saharan Africa, 22% lived in South Asia, 16% lived in East and Southeast Asia, 15% lived in Latin America and the Caribbean, and 12% lived in the Near East and North Africa. The proportions of underweight children included 58% in South Asia, 30% in sub-Saharan Africa, 25% in the Near East and North Africa, 24% in East and Southeast Asia, and 12% in Latin America and the Caribbean. If everyone adopted a vegetarian diet and no food were wasted, there would be enough food to feed 10 billion people. Malnourishment has the harshest effects on children, rural populations, the growing urban poor, and victims of natural disasters.

Schistosomes are parasitic flatworms that infect >200 million people worldwide, causing the chronic, debilitating disease schistosomiasis. Unusual among parasitic helminths, the long-lived adult worms, continuously bathed in blood, take up nutrients directly across the body surface and also by ingestion of blood into the gut. Recent proteomic analyses of the body surface revealed the presence of hydrolytic enzymes, solute, and ion transporters, thus emphasising its metabolic credentials. Furthermore, definition of the molecular mechanisms for the uptake of selected metabolites (glucose, certain amino acids, and water) establishes it as a vital site of nutrient acquisition. Nevertheless, the amount of blood ingested into the gut per day is considerable: for males ∼100 nl; for the more actively feeding females ∼900 nl, >4 times body volume. Ingested erythrocytes are lysed as they pass through the specialized esophagus, while leucocytes become tethered and disabled there. Proteomics and transcriptomics have revealed, in addition to gut proteases, an amino acid transporter in gut tissue and other hydrolases, ion, and lipid transporters in the lumen, implicating the gut as the site for acquisition of essential lipids and inorganic ions. The surface is the principal entry route for glucose, whereas the gut dominates amino acid acquisition, especially in females. Heme, a potentially toxic hemoglobin degradation product, accumulates in the gut and, since schistosomes lack an anus, must be expelled by the poorly understood process of regurgitation. Here we place the new observations on the proteome of body surface and gut, and the entry of different nutrient classes into schistosomes, into the context of older studies on worm composition and metabolism. We suggest that the balance between surface and gut in nutrition is determined by the constraints of solute diffusion imposed by differences in male and female worm morphology. Our conclusions have major implications for

Breast-feeding has always been perceived as a contraceptive measure, probably because it is associated with amenorrhea. However, many pregnancies occur during breast-feeding, often quite soon after delivery. A pregnancy occurring 3 months after delivery is considered at risk for both the mother and child. Three conditions are necessary for a 98% contraceptive efficacy of lactation: total amenorrhea; exclusive breast-feeding on demand, both day and night; and occurrence within the first 6 months postpartum. If the three conditions are met, no additional contraceptive method is needed. Beyond 6 months, another method should be used.

... with a combination of breast- and bottle-feeding. Breast-Feeding Your Baby If your baby is diagnosed with ... use too. If your baby needs surgery after breast-feeding has been established, you can pump your breasts ...

As more mothers elect to breast-feed, more concomitant problems in mothers and babies are reported that are thought to contraindicate breast-feeding. Many frequently cited maternal and infant reasons for stopping breast-feeding are not valid. Breast-feeding can usually be maintained if the physician remembers that breast-feeding is important for the baby and mother and not simply another feeding method. ImagesFigure 1 PMID:21229077

Despite the potential for gossypol toxicosis (particularly in pre-ruminants) and risk factors associated with impaired fertility in bulls, cottonseed products offer a safe alternative feed for cattle producers when fed at recommended levels. Beef producers seeking to lower production costs should consider using cotton byproducts in their feeding programs. If carefully incorporated, cotton byproduct feeds can reduce feed costs while maintaining or increasing the level of cattle performance. Cottonseed meal will remain a standard protein supplement for beef cattle throughout the country. Whole cottonseed has much potential for Southern producers near cotton gins if it is purchased in a timely fashion and fed according to recommendations. Cotton gin trash, cottonseed hulls, and cotton textile mill waste also have potential economic benefits, especially to producers located near cotton and cottonseed processing facilities.

A device for increasing the uniformity of solids within a solids fabrication system, such as a direct light fabrication (DLF) system in which gas entrained powders are passed through the focal point of a moving high-power light which fuses the particles in the powder to a surface being built up in layers. The invention provides a feed through interface wherein gas entrained powders input from stationary input lines are coupled to a rotating head of the fabrication system. The invention eliminates the need to provide additional slack in the feed lines to accommodate head rotation, and therefore reduces feed line bending movements which induce non-uniform feeding of gas entrained powder to a rotating head.

Reliable feeding of coal from storage bins to process requires the knowledge of the behavior of coal during flow. The study of the flow of bulk solids was undertaken in the 1950's and led to the development of flow ability testing equipment and of the Mass Flow concept of design for reliable flow. The theory has since been expanded to two-phase, solids-gas system, and has found world wide application in the design of storage and feeding systems.

Clinicians, public health advisors, nutritionists and others have been attempting to increase breastfeeding rates for the last few decades, with varying degrees of success. We need social science researchers to help us understand the role of infant feeding in the family. Some researchers in the area of food and nutrition have found Pierre Bourdieu's theoretical framework helpful. In this editorial, I introduce some of Bourdieu's ideas and suggest researchers interested in infant feeding should consider testing these theories. PMID:21676218

Near-infrared images of the active galaxy NGC 1097, obtained with the NACO adaptive optics instrument on ESO's Very Large Telescope, disclose with unprecedented detail a complex central network of filamentary structure spiralling down to the centre of the galaxy. These observations provide astronomers with new insights on how super-massive black holes lurking inside galaxies get fed. "This is possibly the first time that a detailed view of the channelling process of matter, from the main part of the galaxy down to the very end in the nucleus is released," says Almudena Prieto (Max-Planck Institute, Heidelberg, Germany), lead author of the paper describing these results. Located at a distance of about 45 million light-years in the southern constellation Fornax (the Furnace), NGC 1097 is a relatively bright, barred spiral galaxy seen face-on. At magnitude 9.5, and thus just 25 times fainter than the faintest object that can be seen with the unaided eye, it appears in small telescopes as a bright, circular disc. NGC 1097 is a very moderate example of an Active Galactic Nucleus (AGN), whose emission is thought to arise from matter (gas and stars) falling into oblivion in a central black hole. However, NGC 1097 possesses a comparatively faint nucleus only, and the black hole in its centre must be on a very strict "diet": only a small amount of gas and stars is apparently being swallowed by the black hole at any given moment. Astronomers have been trying to understand for a long time how the matter is "gulped" down towards the black hole. Watching directly the feeding process requires very high spatial resolution at the centre of galaxies. This can be achieved by means of interferometry as was done with the VLTI MIDI instrument on the central parts of another AGN, NGC 1068 (see ESO PR 17/03), or with adaptive optics [1]. Thus, astronomers [2] obtained images of NGC 1097 with the adaptive optics NACO instrument attached to Yepun, the fourth Unit Telescope of ESO's VLT

Breast feeding has been recommended for the dietary treatment of infants with organic acidaemias, but studies documenting clinical experience are still very few. Nine infants, diagnosed with methylmalonic acidaemia (n = 4), propionic acidaemia (n = 1), isovaleric acidaemia (n = 2) and glutaric acidaemia type I (n = 2) were breast fed after diagnosis. The age of the patients was 28.9+/- 13.4 months (mean +/- SD) (range 10-57 months). Eight patients were diagnosed with clinical symptoms and one because of an affected sibling. After the control of acute metabolic problems, an initial period with a measured volume of expressed breast milk was continued with on-demand breast feeding with the addition of a special essential amino acid mixture and energy supplements. Breast feeding was well tolerated in seven infants with good growth, metabolic control and neurological outcome. The duration of breast feeding was 12.3+/- 7.4 months (mean +/- SD) (range 4-24 months) in these patients. Breast feeding was terminated in the patient with propionic acidaemia because of two acute metabolic episodes requiring hospitalization, and could not be continued in one of the patients with isovaleric acidaemia owing to shortage of breast milk. A decrease in the frequency of infections, acute metabolic episodes and hospital admissions was observed in breast-fed infants. Breast feeding of infants with organic acidaemias is feasible with close monitoring of clinical parameters such as growth, development and biochemistry, including amino acids, organic acids and ammonia.

Human milk is the best way to nurture the human infant. By breast-feeding their babies, mothers provide them with the best opportunities to wholly develop their potential, while protecting the infants and themselves from a whole range of diseases in the near future and in the years to come. Even though these benefits are widely known and there is ample scientific evidence on the topic, it seems from published data that Spanish women are not breast-feeding their babies as much and for as long as they should. Less than 90% start breast-feeding, at 1 month there is already an attrition of 30%, at 3 months more than half of the infants are taking artificial milk and by 6 months only 10% continue to breast-feed their infants. Low birth weight, Caesarean section and low study level are among the more significant factors that negatively affect breast-feeding. There is still work to do to improve this situation. Promotion of breast-feeding among the general population, mothers and health professionals is needed.

Although breast milk provides the best nutrition for the infant, breast-feeding has been decreasing with the growth in use of prepared milk formulas. This paper evaluates the available biblical literature dealing with breast-feeding. The research is unique in character, as it combines contemporary medical knowledge with presentation of cases taken from ancient history. We include no commentaries, but refer only to the words of the Bible exactly as written. Biblical texts relating to breast-feeding were examined and the cases of biblical characters who were breast-fed were studied. The great historical figures such as Isaac, Rebecca, Moses, Samuel, and Joash were breast-fed, openly or secretly. If the biological mother was unable to nurse her child, a wet nurse took over the feeding. Breast-feeding was the only way infants could survive at that time. Weaning day was very important and was celebrated. The wet nurse played an important role in family life, in some cases remaining to live with the infant after weaning, through childhood and even into adulthood. In spite of advanced technology that provides artificial infant formulas, contemporary mothers, just like biblical mothers, should be encouraged to breast-feed their infants.

Plant feeding herbivores excrete most of the ingested chlorophyll (Chl) as partly degraded derivatives lacking the phytol side chain and the central magnesium ion. An ecological role of digested and degraded Chls in the interactions between insects, their food plant and other insects has been described recently. To gain more information on common degradation patterns in plant-feeding insects, the orals secretions and frass of five Lepidopteran caterpillars covering generalists and specialists, namely Spodopteralittoralis, Spodoptera eridania, Heliothis virescens, Helicoverpa armigera, Manduca sexta, and, for comparison, of the leaf beetle larva Chrysomela lapponica were analyzed for chlorophyll catabolites. The major degradation products were determined as pheohorbide a/b and pyropheophorbide a/b by using LC-MS, LC-NMR, UV, and fluorescence spectrometry. The compounds were not present in fresh leaves of the food plants (Phaseolus lunatus, Nicotiana tabacum). The catabolite spectrum in generalists and specialists was qualitatively similar and could be attributed to the action of gut proteins and the strongly alkaline milieu in the digestive tract. Due to the anaerobic environment of the larval gut, the tetrapyrrole core of the Chl catabolites was not cleaved. Substantial amounts of Chl a/b metabolites were strongly complexed by a protein in the mid-gut.

Spodoptera frugiperda Smith (Lepidoptera: Noctuidae) is considered as the most important pest of maize in almost all tropical America. In Argentina, the earwig Doru lineare Eschscholtz (Dermaptera: Forficulidae) has been observed preying on S. frugiperda egg masses in corn crops, but no data about its potential role as a biocontrol agent of this pest have been provided. The predation efficiency of D. lineare on newly emerged S. frugiperda larva was evaluated through a laboratory functional response study. D. lineare showed type II functional response to S. frugiperda larval density, and disc equation estimations of searching efficiency and handling time were (a) = 0.374 and (t) = 182.9 s, respectively. Earwig satiation occurred at 39.4 S. frugiperda larvae. PMID:20575739

AbstractThe fall armyworm Spodoptera frugiperda (Lepidoptera; Noctuidae) is a voracious pest of numerous crops of economic importance throughout the New World. In Brazil, its larvae are attacked by several species of parasitoid wasps, making them potential candidate as biological control agents against this pest. A survey of the parasitoid fauna on S. frugiperda in maize crops throughout Brazil reveals two species of Campoletis, which are morphologicaly very similar species. In this paper we combine these data with pictures from the type material of C. sonorensis and C. flavicincta, as well as their descriptions to provide a redescription to Campoletis sonorensis (Cameron, 1886) using for this both morphological characters and DNA Barcoding (Hebert et al., 2003) information, in an attempt to help with the correct identification of the taxa to improve biological control studies.

The fall armyworm Spodoptera frugiperda (Lepidoptera; Noctuidae) is a voracious pest of numerous crops of economic importance throughout the New World. In Brazil, its larvae are attacked by several species of parasitoid wasps, making them potential candidate as biological control agents against this pest. A survey of the parasitoid fauna on S. frugiperda in maize crops throughout Brazil reveals two species of Campoletis, which are morphologicaly very similar species. In this paper we combine these data with pictures from the type material of C. sonorensis and C. flavicincta, as well as their descriptions to provide a redescription to Campoletis sonorensis (Cameron, 1886) using for this both morphological characters and DNA Barcoding (Hebert et al., 2003) information, in an attempt to help with the correct identification of the taxa to improve biological control studies.

Culture supernatant of Bacillus thuringiensis 9816C had high toxicity against Helicoverpa armigera and Spodoptera exigua. However, it lost insecticidal activities after being bathed in boiling water for 5 min. Acrystalliferous mutants of Bt9816C (Bt9816C-NP1 and Bt9816C-NP2) cured of its endogenous plasmids no longer possessed vip3A gene and toxicity. The 89 kD protein which existed in Bt9816C supernatant disappeared in the two mutants' supernatant; nevertheless, the two mutants still exhibited hemolytic and phospholipase C activity as Bt9816C did. The vip3A gene of Bt9816C, vip3Aa18, was cloned and expressed in Escherichia coli BL21. Bioassay demonstrated that the recombinant E. coli had high toxicity against S. exigua. Taken together, it suggested that Vip3A protein was responsible for the toxicity of Bt9816C culture supernatants.

Although many parasites are known to manipulate the behavior of their hosts, the mechanisms underlying such manipulations are largely unknown. Baculoviruses manipulate the behavior of caterpillar hosts by inducing hyperactivity and by inducing climbing behavior leading to death at elevated positions (tree-top disease or Wipfelkrankheit). Whether hyperactivity and tree-top disease are independent manipulative strategies of the virus is unclear. Recently, we demonstrated the involvement of the protein tyrosine phosphatase ( ptp) gene of the baculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV) in the induction of hyperactivity in Spodoptera exigua larvae. Here we show that AcMNPV ptp is not required for tree-top disease, indicating that in S. exigua baculovirus-induced hyperactivity and tree-top disease are independently induced behaviors that are governed by distinct mechanisms.

Podisus nigrispinus (Dallas) (Heteroptera: Pentatomidae) is one of the most common asopine species in the neotropical region and its occurrence was reported in several countries of South and Central America, as an important biological control agent for many crops. This study was carried out to identify the imidacloprid impacts on the functional response of predator P. nigrispinus fed on Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) strain resistant to lambda-cyhalothrin, on Bt cotton expressing Cry1Ac (Bollgard(®)). Spodoptera frugiperda larvae were used in the following conditions: resistant (1) and susceptible (2) strains to lambda-cyhalothrin fed on Bollgard(®) cotton leaves (DP 404 BG); and resistant (3) and susceptible (4) strains to lambda-cyhalothrin fed on non-genetically modified cotton leaves (cultivar DP4049). The predatory behavior of P. nigrispinus was affected by imidacloprid and the type II asymptotic curve was the one that best described the functional response data. Handling time (T h ) of predator females did not differ among treatments in the presence of imidacloprid. The attack rate did decrease, however, due to an increase in the density of larvae offered. Regardless of the treatment (S. frugiperda strain or cotton cultivar), the predation of P. nigrispinus females on S. frugiperda larvae was significantly lower when exposed to imidacloprid, especially at a density of 16 larvae/predator. The predation behavior of P. nigrispinus on S. frugiperda larvae is affected by the insecticide imidacloprid showing that its applications should be used in cotton crop with caution.

Spodoptera frugiperda larvae are highly resistant to oral infection by Autographa californica multiple nucleopolyhedrovirus (AcMNPV) (LD(50), approximately 9200 occlusions), but extremely susceptible to budded virus within the haemocoel (LD(50), <1 p.f.u.). The inability of AcMNPV occlusion-derived virus (ODV) to establish primary infections readily within midgut cells accounts for a major proportion of oral resistance. To determine whether inappropriate binding of AcMNPV ODV to S. frugiperda midgut cells contributes to lack of oral infectivity, the binding and fusion properties of AcMNPV ODV were compared with those of the ODV of a new isolate of Spodoptera frugiperda multiple nucleopolyhedrovirus (SfMNPV) obtained from a field-collected larva (oral LD(50), 12 occlusions). By using a fluorescence-dequenching assay conducted in vivo, it was found that AcMNPV ODV bound to the midgut epithelia of S. frugiperda larvae at approximately 15 % of the level of SfMNPV ODV, but that, once bound, the efficiencies of fusion for the two ODVs were similar: 60 % for AcMNPV and 53 % for SfMNPV. Whilst the difference in binding efficiencies was significant, it could not account entirely for the observed differences in infectivity. Competition experiments, however, revealed that, in S. frugiperda larvae, SfMNPV ODV bound to a midgut cell receptor that was not bound by AcMNPV ODV, indicating that ODV interaction with a specific receptor(s) was necessary for productive infection of midgut columnar epithelial cells. Fusion in the absence of this ligand-receptor interaction did not result in productive infections.

ML (MD2-related lipid recognition) proteins are known to enhance innate immune responses in mammals. This study reports the analysis of the putative ML gene family in Arabidopsis thaliana and suggests a role for the ML3 gene in herbivory-associated responses in plants. Feeding by larvae of the Lepidopteran generalist herbivore Spodopteralittoralis and larvae of the specialist herbivore Plutella xylostella activated ML3 transcription in leaf tissues. ML3 loss-of-function Arabidopsis plants were compromised in the upregulation of herbivory-induced genes and displayed a semi-dwarf phenotype. Herbivory bioassays showed that larvae of S. littoralis fed on ml3 mutant plants gained more weight compared to larvae fed on wild-type plants while larvae of P. xylostella did not show any significant difference. Virus-induced gene silencing of ML3 expression in plants compromised in jasmonic acid (JA) and salicylic acid (SA) signalling revealed a complex role of ML3 in JA/defence signalling affecting both JA- and SA-dependent responses. The data suggest that ML3 is involved in herbivory-mediated responses in Arabidopsis and that it has a potential role in herbivory-associated molecular pattern recognition.

• Herbivore-induced systemic resistance occurs in many plants and is commonly assumed to be adaptive. The mechanisms triggered by leaf-herbivores that lead to systemic resistance are largely understood, but it remains unknown how and why root herbivory also increases resistance in leaves. • To resolve this, we investigated the mechanism by which the root herbivore Diabrotica virgifera induces resistance against lepidopteran herbivores in the leaves of Zea mays. • Diabrotica virgifera infested plants suffered less aboveground herbivory in the field and showed reduced growth of Spodopteralittoralis caterpillars in the laboratory. Root herbivory did not lead to a jasmonate-dependent response in the leaves, but specifically triggered water loss and abscisic acid (ABA) accumulation. The induction of ABA by itself was partly responsible for the induction of leaf defenses, but not for the resistance against S. littoralis. Root-herbivore induced hydraulic changes in the leaves, however, were crucial for the increase in insect resistance. • We conclude that the induced leaf resistance after root feeding is the result of hydraulic changes, which reduce the quality of the leaves for chewing herbivores. This finding calls into question whether root-herbivore induced leaf-resistance is an evolved response.

Introduction The nemertean pilidium is a long-lived feeding larva unique to the life cycle of a single monophyletic group, the Pilidiophora, which is characterized by this innovation. That the pilidium feeds on small planktonic unicells seems clear; how it does so is unknown and not readily inferred, because it shares little morphological similarity with other planktotrophic larvae. Results Using high-speed video of trapped lab-reared pilidia of Micrura alaskensis, we documented a multi-stage feeding mechanism. First, the external ciliation of the pilidium creates a swimming and feeding current which carries suspended prey past the primary ciliated band spanning the posterior margins of the larval body. Next, the larva detects prey that pass within reach, then conducts rapid and coordinated deformations of the larval body to re-direct passing cells and surrounding water into a vestibular space between the lappets, isolated from external currents but not quite inside the larva. Once a prey cell is thus captured, internal ciliary bands arranged within this vestibule prevent prey escape. Finally, captured cells are transported by currents within a buccal funnel toward the stomach entrance. Remarkably, we observed that the prey of choice – various cultured cryptomonads – attempt to escape their fate. Conclusions The feeding mechanism deployed by the pilidium larva coordinates local control of cilia-driven water transport with sensorimotor behavior, in a manner clearly distinct from any other well-studied larval feeding mechanisms. We hypothesize that the pilidium’s feeding strategy may be adapted to counter escape responses such as those deployed by cryptomonads, and speculate that similar needs may underlie convergences among disparate planktotrophic larval forms. PMID:23927417

The apparatus of the present invention is a melter housing having a pretreat chamber heated with a feed material heater that is partially isolated from a melter chamber. The method of the present invention has the steps of introducing feed material into a pretreat chamber and heating the feed material to a softening temperature of the feed material, and passing the pretreated feed material to a melter chamber.

Managing infants and children with difficult feeding problems, such as intractable vomiting and dysmotility, can be challenging. Maintaining sufficient enteral feeding is the ultimate goal. However, the options for establishing this goal may be limited. One option is gastrojejunostomy tube feeding, a technique developed in 1984. This review discusses indications and limitations of gastrojejunostomy tube feeding in children. It also discusses alternative options for gastrojejunostomy tube feeding in view of the available evidence.

Beer consumption by nursing women altered the sensory qualities of their milk and the behavior of their infants during breast-feeding in the short term. The infants consumed significantly less milk during the 4-hr testing sessions in which their mothers drank alcoholic beer compared to when the mothers drank nonalcoholic beer; this decrease in milk intake was not due to a decrease in the number of times the babies fed. Although the infants consumed less of the alcohol-flavored milk, the mothers believed their infants had ingested enough milk, reported that they experienced a letdown during nursing, and felt they had milk remaining in their breasts at the end of the majority of feedings. Moreover, the mothers terminated the feeds the same percentage of time on both testing days. The mechanism by which the consumption of alcoholic beer by lactating women decreases milk intake by their nurslings remains to be determined. PMID:8293892

Beer consumption by nursing women altered the sensory qualities of their milk and the behavior of their infants during breast-feeding in the short term. The infants consumed significantly less milk during the 4-hr testing sessions in which their mothers drank alcoholic beer compared to when the mothers drank nonalcoholic beer; this decrease in milk intake was not due to a decrease in the number of times the babies fed. Although the infants consumed less of the alcohol-flavored milk, the mothers believed their infants had ingested enough milk, reported that they experienced a letdown during nursing, and felt they had milk remaining in their breasts at the end of the majority of feedings. Moreover, the mothers terminated the feeds the same percentage of time on both testing days. The mechanism by which the consumption of alcoholic beer by lactating women decreases milk intake by their nurslings remains to be determined.

An advanced static feed water electrolysis module (SFWEM) and associated instrumentation for generating breathable O2 was developed. The system also generates a H2 byproduct for use in an air revitalization system for O2 recovery from metabolic CO2. Special attention was given to: (1) eliminating water feed compartment degassing, (2) eliminating need for zero gravity condenser/separators, (3) increasing current density capability, and (4) providing a self contained module so that operation is independent of laboratory instrumentation and complicated startup/shutdown procedures.

Plants produce a wide variety of defensive metabolites to protect themselves against herbivores and pathogens. Non-protein amino acids, which are present in many plant species, can have a defensive function through their mis-incorporation during protein synthesis and/or inhibition of biosynthetic pathways in primary metabolism. 5-Hydroxynorvaline was identified in a targeted search for previously unknown non-protein amino acids in the leaves of maize (Zea mays) inbred line B73. Accumulation of this compound increases during herbivory by aphids (Rhopalosiphum maidis, corn leaf aphid) and caterpillars (Spodoptera exigua, beet armyworm), as well as in response to treatment with the plant signalling molecules methyl jasmonate, salicylic acid and abscisic acid. In contrast, ethylene signalling reduced 5-hydroxynorvaline abundance. Drought stress induced 5-hydroxynorvaline accumulation to a higher level than insect feeding or treatment with defence signalling molecules. In field-grown plants, the 5-hydroxynorvaline concentration was highest in above-ground vegetative tissue, but it was also detectable in roots and dry seeds. When 5-hydroxynorvaline was added to aphid artificial diet at concentrations similar to those found in maize leaves and stems, R. maidis reproduction was reduced, indicating that this maize metabolite may have a defensive function. Among 27 tested maize inbred lines there was a greater than 10-fold range in the accumulation of foliar 5-hydroxynorvaline. Genetic mapping populations derived from a subset of these inbred lines were used to map quantitative trait loci for 5-hydroxynorvaline accumulation to maize chromosomes 5 and 7.

Plants produce a wide variety of defensive metabolites to protect themselves against herbivores and pathogens. Non-protein amino acids, which are present in many plant species, can have a defensive function through their mis-incorporation during protein synthesis and/or inhibition of biosynthetic pathways in primary metabolism. 5-Hydroxynorvaline was identified in a targeted search for previously unknown non-protein amino acids in the leaves of maize (Zea mays) inbred line B73. Accumulation of this compound increases during herbivory by aphids (Rhopalosiphum maidis, corn leaf aphid) and caterpillars (Spodoptera exigua, beet armyworm), as well as in response to treatment with the plant signalling molecules methyl jasmonate, salicylic acid and abscisic acid. In contrast, ethylene signalling reduced 5-hydroxynorvaline abundance. Drought stress induced 5-hydroxynorvaline accumulation to a higher level than insect feeding or treatment with defence signalling molecules. In field-grown plants, the 5-hydroxynorvaline concentration was highest in above-ground vegetative tissue, but it was also detectable in roots and dry seeds. When 5-hydroxynorvaline was added to aphid artificial diet at concentrations similar to those found in maize leaves and stems, R. maidis reproduction was reduced, indicating that this maize metabolite may have a defensive function. Among 27 tested maize inbred lines there was a greater than 10-fold range in the accumulation of foliar 5-hydroxynorvaline. Genetic mapping populations derived from a subset of these inbred lines were used to map quantitative trait loci for 5-hydroxynorvaline accumulation to maize chromosomes 5 and 7. PMID:25271262

Integrin is a cell-surface protein consisting of α and β heterodimers. A predicted amino acid sequence of an integrin subunit of the diamondback moth, Plutella xylostella, was highly homologous to other lepidopteran β1 subunits and possessed essential functional domains. The β1 integrin of P. xylostella (βPx1) was expressed in all developmental stages of P. xylostella. It was also expressed in all tested tissues including hemocyte, fat body, gut, and epidermis of last instar. When βPx1 expression was suppressed by injection of dsRNA specific to βPx1 (dsRNA(βPx1)), the treated larvae exhibited significant suppression in immune response and also suffered significant larval mortality. When dsRNA(βPx1) was orally fed to young larvae, it suppressed the expression of âPx1 and resulted in a significant mortality. By contrast, a dsRNA specific to β1 subunit of Spodoptera exigua gave little adverse effects on βPx1 expression and larval development when it was treated by injection or oral administration, though these two genes showed 71% sequence homology. These results suggest a target-specific RNA interference of dsRNA(βPx1), which causes significant mortality to P. xylostella by feeding treatment.

Some specialist insects feed on plants rich in secondary compounds, which pose a major selective pressure on both the phytophagous and the gut microbiota. However, microbial communities of toxic plant feeders are still poorly characterized. Here, we show the bacterial communities of the gut of two specialized Lepidoptera, Hyles euphorbiae and Brithys crini, which exclusively feed on latex-rich Euphorbia sp. and alkaloid-rich Pancratium maritimum, respectively. A metagenomic analysis based on high-throughput sequencing of the 16S rRNA gene revealed that the gut microbiota of both insects is dominated by the phylum Firmicutes, and especially by the common gut inhabitant Enterococcus sp. Staphylococcus sp. are also found in H. euphorbiae though to a lesser extent. By scanning electron microscopy, we found a dense ring-shaped bacterial biofilm in the hindgut of H. euphorbiae, and identified the most prominent bacterium in the biofilm as Enterococcus casseliflavus through molecular techniques. Interestingly, this species has previously been reported to contribute to the immobilization of latex-like molecules in the larvae of Spodoptera litura, a highly polyphagous lepidopteran. The E. casseliflavus strain was isolated from the gut and its ability to tolerate natural latex was tested under laboratory conditions. This fact, along with the identification of less frequent bacterial species able to degrade alkaloids and/or latex, suggest a putative role of bacterial communities in the tolerance of specialized insects to their toxic diet. PMID:27446044

Two studies examined the effects of breast-feeding on maternal cardiovascular function. In the first experiment, groups of breast-feeding and bottle-feeding women were compared on preejection period (PEP), heart rate (HR), cardiac output (CO), and total peripheral resistance (TPR) recorded for 1-min periods before and during standard laboratory stressors. Compared with bottle-feeders, breast-feeders had higher CO throughout the session, and greater decreases in CO and increases in TPR during cold pressor. In the second experiment, HR and blood pressure (BP) were compared before and after one breast-feeding and one bottle-feeding session in a within-subjects design. Both feeding methods increased BP but decreased HR, and systolic BP was higher for the breast-feeding than the bottle-feeding condition. Both studies support the notion that breast-feeding alters maternal cardiovascular function, possibly through the actions of oxytocin.

Most commercially cultivated fish are raised in high-density culture systems where the assumption is that the contribution of natural foods to the nutrition of the fish is insignificant. Thus, intensively cultured fish must be fed a nutritionally complete feed. A short section on the concept and im...

Non-feed uses for alfalfa such as biomass energy and phytoremediation could increase alfalfa acreage and improve farm profitability. The new bio-energy alfalfa and production system increased forage yield and ethanol production. New alfalfas with enhanced nitrogen cycling capacities would protect wa...

One of the greatest advantages of feeding exclusively breast-milk is the continuous provision of immunoglobulin A, especially during the first days of life, and of leucocytes with macrophage function as well as unspecific, antiinfectious agents like lactoferrin, lysozyme and neuraminic acid. It seems, that the organism is protected against allergic reactions at the mucosa level of the small intestine caused by the penetration of "foreign" protein by feeding exclusively breast-milk especially during the first weeks and months of life. During the first months of the infant's life an increased supply of iron results from the higher content of iron in breast-milk as compared to cow's milk, and the better absorption of the iron from breast-milk. Just because of this (the better provision with iron from natural food) solid foods should not be added to the infant's diet before 6 months of age. One of the disadvantages of breast-feeding is the passage of unwanted substances from breast-milk to the infant. First of all the chlorinated hydrocarbons have to be mentioned within this context. However, a decreasing tendency can be assumed according to recent investigations. An increasing tendency in breast-milk, though not confirmed, seems possible only for the polychlorinated biphenyls. Nevertheless, for the pediatrician no reason to advise against breast-feeding results from the unwanted admixtures of chlorinated hydrocarbons in breast-milk. One should rather vigorously propagate to feed as many children as possible exclusively with breast-milk over a period of 4 to 6 months.

Homologous and heterologous nucleopolyhedroviruses (NPVs) were assayed to determine the most effective NPV against beet armyworm larvae, Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae)(SeMNPV). Included were three isolates from S. exigua, one isolate each from S. littoralis Boisduval, S. litura...

A feeding mechanism and method for feeding minute items, such as capacitors, resistors, or solder preforms. The mechanism is adapted to receive a plurality of the randomly-positioned and randomly-oriented extremely small or minute items, and to isolate, orient, and position one or more of the items in a specific repeatable pickup location wherefrom they may be removed for use by, for example, a computer-controlled automated assembly machine. The mechanism comprises a sliding shelf adapted to receive and support the items; a wiper arm adapted to achieve a single even layer of the items; and a pushing arm adapted to push the items into the pickup location. The mechanism can be adapted for providing the items with a more exact orientation, and can also be adapted for use in a liquid environment.

A feeding mechanism and method for feeding minute items, such as capacitors, resistors, or solder preforms. The mechanism is adapted to receive a plurality of the randomly-positioned and randomly-oriented extremely small or minute items, and to isolate, orient, and position the items in a specific repeatable pickup location wherefrom they may be removed for use by, for example, a computer-controlled automated assembly machine. The mechanism comprises a sliding shelf adapted to receive and support the items; a wiper arm adapted to achieve a single even layer of the items; and a pushing arm adapted to push the items into the pickup location. The mechanism can be adapted for providing the items with a more exact orientation, and can also be adapted for use in a liquid environment.

This plan documents the Waste Feed Delivery Program test and evaluation planning and implementation approach. The purpose of this document is to define and communicate the Waste Feed Delivery Program Test and Evaluation scope, objectives, planning and implementation approach.

... 000806.htm Infant Formula – Buying, Preparing, Storing, and Feeding To use the sharing features on this page, ... brush to get at hard-to-reach places. Feeding Formula to Baby Here is a guide to ...

Hydrodynamics plays a dominant role for small planktonic flagellates and shapes their survival strategies. The high diversity of beat patterns and arrangements of appendages indicates different strategies balancing the trade-offs between the general goals, i.e., energy-efficient swimming, feeding, and predator avoidance. One type of flagellated algae that we observe, are haptophytes, which possess two flagella for flow creation and one so-called haptonema, a long, rigid structure fixed on the cell body, which is used for prey capture. We present videos and flow fields obtained using velocimetry methods around freely swimming haptophytes and other flagellates, which we compare to analytical results obtained from point force models. The observed and modelled flows are used to analyse how different morphologies and beat patterns relate to different feeding or swimming strategies, such as the capture mechanism in haptophytes. The Centre for Ocean Life is a VKR center of excellence supported by the Villum foundation.

Suspension-feeding bivalve molluscs are confronted with a wide range of materials in the benthic marine environment. These materials include various sized plankton and the organic material derived from it, macroalgae, detritus and a diversity of microbial parasites that have adapted life stages to survive in the water column. For bivalve parasites to infect hosts though, they must first survive and remain infectious in the water column to make initial contact with hosts, and once in contact, enter and overcome elaborate pathways for particle sorting and selection. Even past these defenses, bivalve parasites are challenged with efficient systems of mechanical and chemical digestion and highly evolved systems of innate immunity. Here we review how bivalve parasites evade these hurdles to complete their life cycles and establish within bivalve hosts. We broadly cover significant viral, bacterial, and protozoan parasites of marine bivalve molluscs, and illustrate the emergent properties of these host-parasite systems where parasite transmission occurs through suspension feeding.

There is increasing evidence to suggest that the cannabinoid system is a crucial mechanism in the regulation of feeding and metabolism. It is against this background that a cannabinoid antagonist, rimonabant, is about to come onto the market for the treatment of obesity. Moreover, in addition to weight-loss effect, this drug has a beneficial effect on the so-called metabolic syndrome, with changes in the lipid and glucidic metabolism not observed for other anti-obesity drugs currently available. We present a review of current knowledge in this field and data from our own studies: genetic studies of this system in eating disorders and in obesity and studies of localization of cannabinoid receptors at sites related to feeding. These studies support a state of cannabinoid hyperactivity in obesity; furthermore, such hyperactivity can constitute a prognostic factor.

Evolutionary transitions between aquatic and terrestrial environments are common in vertebrate evolution. These transitions require major changes in most physiological functions, including feeding. Emydid turtles are ancestrally aquatic, with most species naturally feeding only in water, but some terrestrial species can modulate their feeding behavior appropriately for both media. In addition, many aquatic species can be induced to feed terrestrially. A comparison of feeding in both aquatic and terrestrial environments presents an excellent opportunity to investigate the evolution of terrestrial feeding from aquatic feeding, as well as a system within which to develop methods for studying major evolutionary transitions between environments. Individuals from eight species of emydid turtles (six aquatic, two terrestrial) were filmed while feeding underwater and on land. Bite kinematics were analyzed to determine whether aquatic turtles modulated their feeding behavior in a consistent and appropriate manner between environments. Aquatic turtles showed consistent changes between environments, taking longer bites and using more extensive motions of the jaw and hyoid when feeding on land. However, these motions differ from those shown by species that naturally feed in both environments and mostly do not seem to be appropriate for terrestrial feeding. For example, more extensive motions of the hyoid are only effective during underwater suction feeding. Emydids evolving to feed on land probably would have needed to evolve or learn to overcome many, but not all, aspects of the intrinsic emydid response to terrestrial feeding. Studies that investigate major evolutionary transitions must determine what responses to the new environment are shown by naïve individuals in order to fully understand the evolutionary patterns and processes associated with these transitions.

The article provides an overview of the available literature on problems of infant nutrition, and shows the historical development of the principles of infant feeding. It discusses in greater detail the use of goat milk as a basis for infant nutrition. It notes the need for a comparative analysis of breast milk substitutes, and for clinical studies evaluating the value of goat milk in infant nutrition.

Black holes are the ultimate prisons of the Universe, regions of spacetime where the enormous gravity prohibits matter or even light to escape to infinity. Yet, matter falling toward the black holes may shine spectacularly, generating the strongest source of radiation. These sources provide us with astrophysical laboratories of extreme physical conditions that cannot be realized on Earth. This chapter offers a review of the basic menus for feeding matter onto black holes and discusses their observational implications.

When a child is born with a feeding and/or swallowing problem, it affects the entire family. Parents, mothers specifically, internalize the problem and come to believe it is due to something they've done. Families are put on an emotional rollercoaster with the ups and downs being dependant on how much the child has eaten that day. In this article,…

A fluidized bed boiler feed system for the combustion of pulverized coal. Coal is first screened to separate large from small particles. Large particles of coal are fed directly to the top of the fluidized bed while fine particles are first mixed with recycled char, preheated, and then fed into the interior of the fluidized bed to promote char burnout and to avoid elutriation and carryover.

A critical point of nasogastric feeding tube placement, potentially resulting in an unsafe and/or non-effective operation of the device, is the monitoring of its proper placement into the stomach. A properly obtained and interpreted radiograph is currently recommended to confirm placement. We reported the case of a 68-year-old demented woman referred for complicated dysphagia. A nasogastric tube was blindly inserted and its placement was confirmed by the radiologist. Enteral nutrition was initiated but the patient began to vomit immediately. After reviewing the radiograph it was understood that a gastric loop in the tube and its tip pointing upwards did not allow a safe infusion of the feeding formula. It is not enough having the radiologist reporting that a nasogastric feeding tube is placed in the stomach; the inclusion in the report of specific warnings on any potential cause of malfunctioning of the device should be considered. The presence of a gastric loop should be taken into account as a cause of potential malfunctioning.

Published studies of consumer feeding preferences using foods that experience autogenic change in mass, numbers, area, etc., on the time scale of a feeding trial fail to employ appropriate statistical analyses to incorporate controls for those food changes occurring in the absence of the consumer. The studies that run controls typically use them to calculate a constant "correction factor", which is subtracted prior to formal data analysis. This procedure constitutes a non-rigorous suppression of variance that overstates the statistical significance of observed differences. The appropriate statistical analysis for preference tests with two foods is usually a simple t-test performed on the between-food differences in loss of mass (or numbers, area, etc.) comparing the results of experimentals with consumers to controls without consumers. Application of this recommended test procedure to an actual data set illustrates how low replication in controls, which is typical of most studies of feeding preference, inhibits detection of an apparently large influence of previous mechanical damage (simulated grazing) in reducing the attractiveness of a brown alga to a sea urchin.

Children with genetic syndromes frequently have feeding problems and swallowing dysfunction as a result of the complex interactions between anatomical, medical, physiological, and behavioral factors. Feeding problems associated with genetic disorders may also cause feeding to be unpleasant, negative, or even painful because of choking, coughing,…

Whenever possible, oral feeding is the preferred method in neonatal feeding. However, many premature infants are unable to suck and swallow effectively; in these cases alternative methods of nutrient delivery must be used. We briefly review the different feeding methods used in neonatal units, with particular attention to their theoretical advantages, disadvantages and to the current best evidence available.

Researchers use the Caregiver's Feeding Styles Questionnaire (CFSQ) to categorize parent feeding into authoritative, authoritarian, indulgent, and uninvolved styles. The CFSQ assesses self-reported feeding and classifies parents using median splits which are used in a substantial body of parenting l...

Applies principles of the transdisciplinary team approach to the needs of children with feeding disorders and describes a pediatric feeding team at the University of South Alabama Children's and Women's Hospital. Case studies illustrate the principles. Advantages and challenges in the transdisciplinary approach to feeding disorders are discussed.…

Infant feeding decisions are some of the most important choices parents make. Breast milk or formula is the first decision made in infant feeding. Complementary feeding is common among very young children in Turkey. Therefore, the aim of this research is to focus on the introduction of solid foods, and to determine the relationship between…

Food availability is a primary driver of avian population regulation. However, few studies have considered the effects of what is essentially a massive supplementary feeding experiment: the practice of wild bird feeding. Bird feeding has been posited as an important factor influencing the structure of bird communities, especially in urban areas, although experimental evidence to support this is almost entirely lacking. We carried out an 18-mo experimental feeding study at 23 residential properties to investigate the effects of bird feeding on local urban avian assemblages. Our feeding regime was based on predominant urban feeding practices in our region. We used monthly bird surveys to compare avian community composition, species richness, and the densities of local species at feeding and nonfeeding properties. Avian community structure diverged at feeding properties and five of the commonest garden bird species were affected by the experimental feeding regime. Introduced birds particularly benefitted, with dramatic increases observed in the abundances of house sparrow (Passer domesticus) and spotted dove (Streptopelia chinensis) in particular. We also found evidence of a negative effect on the abundance of a native insectivore, the grey warbler (Gerygone igata). Almost all of the observed changes did not persist once feeding had ceased. Our study directly demonstrates that the human pastime of bird feeding substantially contributes to the structure of avian community in urban areas, potentially altering the balance between native and introduced species.

The objective of this study was to determine the obstetric care providers' roles in breast-feeding promotion during prenatal care. A questionnaire addressing breast-feeding issues was sent to family practitioners (FP), obstetric-gynecologists (OB/GYN), and nurse midwives (NM) in Iowa, USA. All NM, 97% of FP, and 85% of OB/GYN reported asking infant feeding preference-usually only at the first prenatal visit. NM (73%) were most likely to provide extensive breast-feeding counseling. OB/GYN (68%) and FP physicians (90%) reported doing their own breast-feeding counseling. Breast examinations targeting future breast-feeding problems were done in 82% to 84% of patients. NM practices shared more information supportive of breast-feeding. Nearly all providers offered prenatal classes, but only 41% of FP offered breast-feeding classes. Free formula samples were available in 73% of FP, 54% of OB/GYN, and 36% NM offices. Pamphlets on formula feeding and also breast-feeding were readily available. Overall NM (64%) reported being strong breast-feeding advocates compared to only 13% of FP and 7% of OB/GYN. In conclusion, little promotion of breast-feeding occurs in most prenatal practice settings.

Adult stable flies, (Stomoxys calcitrans (L.)), are known to feed readily on sugars in the laboratory. However, little is known concerning the extent of stable fly sugar feeding in wild populations. We examined the frequency of sugar feeding in stable flies in rural and urban environments. In additi...

A mother's nutritional status during pregnancy has important implications for both her own health and her ability to produce and breast-feed a healthy infant. Knowledge about adequate maternal nutrition during pregnancy is incomplete, however, and there is still considerable debate about the level of extra energy needed by a pregnant woman. A woman's usual nutritional requirements increase during pregnancy to meet her needs and those of the growing fetus. Additional energy is needed because of increased basal metabolism, the greater cost of physical activity, and the normal accumulation of fat as the energy reserve. The protein, vitamin and mineral requirements of the mother also increase during pregnancy, but the precise amounts for the last two are still a matter for discussion. A woman's weight increments during pregnancy vary between privileged and underprivileged communities. In addition to calcium, phosphorus and iron, a mother provides considerable amounts of protein and fat for fetal growth. Placental metabolism and placental blood flow, which are interrelated, are the most critical factors for fetal development.The nutritional requirements of healthy newborns vary widely according to their weight, gestational age, rate of growth, as well as environmental factors. However, recommendations for some components may be derived from the average composition of early human milk and the amounts consumed by healthy, mature newborns who are following a normal postpartum clinical course. The water requirements of infants are related to their caloric consumption, activity, rate of growth, and the ambient temperature. A postnatal weight loss of 5-8% of body weight is usual during the first few days of life in mature newborn infants; in contrast, infants who experienced intrauterine malnutrition lose little or no weight at all.The dynamic process of mother-newborn interaction from the first hours of life is intimately related to successful early breast-feeding. If this

This manual provides guidelines for safe feeding practices for students with disabilities in Oregon schools. Stressed is the importance of distinguishing between feeding for the maintenance of health and feeding for the acquisition of skills. Individual sections cover: definitions of feeding programs; the school district responsibility; risks;…

Little is known about the feeding behaviors and problems with feeding in toddlers. In the present questionnaire study, data were collected on the feeding behaviors and feeding problems in a relatively large (n = 422) sample of Dutch healthy toddlers (i.e. 18-36 months old) who lived at home with their parents. Results show that three meals a day…

Deregulated reactive oxygen species (ROS) production can lead to the disruption of structural and functional integrity of cells as a consequence of reactive interaction between ROS and various biological components. Catalase (CAT) is a common enzyme existing in nearly all organisms exposed to oxygen, which decomposes harmful hydrogen peroxide, into water and oxygen. In this study, the full length sequence that encodes CAT-like protein from Spodoptera litura named siltCAT (GenBank accession number: JQ_663444) was cloned and characterized. Amino acid sequence alignment showed siltCAT shared relatively high conservation with other insect, especially the conserved residues which defined heme and NADPH orientation. Expression pattern analysis showed that siltCAT mRNA was mainly expressed in the fat body, midgut, cuticle and malpighian tube, and as well as over last instar larvae, pupa and adult stages. RNA interference was used to silence CAT gene in SL-1 cells and the fourth-instar stage of S. litura larvae respectively. Our results provided evidence that CAT knockdown induced ROS generation, cell cycle arrest and apoptosis in SL-1 cells. It also confirmed the decrease in survival rate because of increased ROS production in experimental groups injected with double-stranded RNA of CAT (dsCAT). This study implied that ROS scavenging by CAT is important for S. litura survival. PMID:23555693

The effects of different application technologies were evaluated on the concentration, viability, and efficiency of infective juveniles of the nematodes Heterorhabditis indica Poinar, Karunakar & David and Steinernema sp. (IBCB-n6) to control Spodoptera frugiperda Smith on corn plants. Two hundred and eighty infective juveniles of Steinernema sp. were required to kill 100% third-instar fall armyworms in petri dishes, as compared to 400 infective juveniles of the H. indica nematode to obtain 75% fall armyworm control. It is possible to spray entomopathogenic nematodes without significant loss in their concentration and viability, with equipment that produces electrical charges to the spraying mix, and with those using hydraulic and rotary nozzle tips. The concentrations of infective juveniles of H. indica and Steinernema sp. nematodes were reduced by 28% and 53%, respectively, when hydraulic spraying nozzles that require 100-mesh filtrating elements were used. Tensoactive agents of the organosilicone and ethoxylate groups did not affect the viability of infective juveniles of Steinernema sp. juveniles. Spraying corn plants (V6 growth stage) with up to 288 million infective juveniles of Steinernema sp. per hectare, diluted in the spraying mix up to 800 L ha-1, with 0.01% ethoxylate tensoactive agent, or at the same volume followed by artificial rain (6 mm water depth) was not sufficient to control S. frugiperda in a controlled environment.

A novel flavonoid, quercetin, was isolated from the medicinal plant Euphorbia hirta L. through chromatography techniques including: TLC, Column chromatography, NMR and then screened for toxicity to larvae of Spodoptera litura Fab. Bioassays were used to analyze pupal weight, survival rate, fecundity, egg hatchability, population growth index, Nutritional index and histopathology of treated larvae at a range of E. hirta extract concentrations. Results of toxicity assays demonstrated that, 6 ppm of quercetin caused 94.6% mortality of second, 91.8% of third, 88% of fourth, and 85.2% of fifth instars respectively. The lethal concentrations (LC50 and LC90) was calculated as 10.88 and 69.91 ppm for fourth instar larvae. The changes in consumption ratio and approximate digestibility produced a reduction in growth rates. Histopathology examinations revealed that the cell organelles were severely infected. Analyses of earthworm toxicity effects resulted in significantly lower rates compared to synthetic insecticides (chloropyrifos and cypermethrin). These results suggests that the botanical compound (quercetin), could have a part as a new biorational product which provides an ecofriendly alternative. Validation of the potential of quercetin, still needs to be demonstrated under field conditions, where formulation will be important in maintaining the activity.

Many parasites enhance their dispersal and transmission by manipulating host behaviour. One intriguing example concerns baculoviruses that induce hyperactivity and tree-top disease (i.e., climbing to elevated positions prior to death) in their caterpillar hosts. Little is known about the underlying mechanisms of such parasite-induced behavioural changes. Here, we studied the role of the ecdysteroid UDP-glucosyltransferase (egt) gene of Spodoptera exigua multiple nucleopolyhedrovirus (SeMNPV) in tree-top disease in S. exigua larvae. Larvae infected with a mutant virus lacking the egt gene exhibited a shorter time to death and died before the induction of tree-top disease. Moreover, deletion of either the open reading frame or the ATG start codon of the egt gene prevented tree-top disease, indicating that the EGT protein is involved in this process. We hypothesize that SeMNPV EGT facilitates tree-top disease in S. exigua larvae by prolonging the larval time to death. Additionally, we discuss the role of egt in baculovirus-induced tree-top disease.

Analyses of extracts of pheromone glands and of volatiles from calling female fall armyworm moths,Spodoptera frugiperda (J.E. Smith), revealed the presence of the following compounds: dodecan-1-ol acetate, (Z)-7-dodecen-1-ol acetate, 11-dodecen-1-ol acetate, (Z)-9-tetradecenal, (Z)-9-tetradecen-1-ol acetate, (Z)-11-hexadecenal, and (Z)-11-hexadecen-1-ol acetate. The volatiles emitted by calling females differed from the gland extract in that the two aldehydes were absent. Field tests were conducted with sticky traps baited with rubber septa formulated to release blends with the same component ratios as those emitted by calling females. These tests demonstrated that both (Z)-7-dodecen-1-ol acetate and (Z)-9-tetradecen-1-ol acetate are required for optimum activity and that this blend is a significantly better lure than either virgin females or 25 mg of (Z)-9-dodecen-1-ol acetate in a polyethylene vial, the previously used standard. Addition of the other three acetates found in the volatiles did not significantly increase the effectiveness of the two-component blend as a bait for Pherocon 1C or International Pheromones moth traps.

Vip proteins, a new group of insecticidal toxins produced by Bacillus thuringiensis, are effective against specific pests including Spodoptera litura. Here, we report construction of a transcriptome database of S. litura by de novo assembly along with detection of the transcriptional response of S. litura larvae to Vip3Aa toxin. In total, 56,498 unigenes with an N50 value of 1,853 bp were obtained. Results of transcriptome abundance showed that Vip3Aa toxin provoked a wide transcriptional response of the S. litura midgut. The differentially expressed genes were enriched for immunity-related, metabolic-related and Bt-related genes. Twenty-nine immunity-related genes, 102 metabolic-related genes and 62 Bt-related genes with differential expression were found. On the basis of transcriptional profiling analysis, we focus on the functional validation of trypsin which potentially participated in the activation of Vip3Aa protoxin. Zymogram analysis indicated that the presence of many proteases, including trypsin, in S. litura larvae midgut. Results of enzymolysis in vitro of Vip3Aa by trypsin, and bioassay and histopathology of the trypsin-digested Vip3Aa toxin showed that trypsin was possibly involved in the Vip3Aa activation. This study provides a transcriptome foundation for the identification and functional validation of the differentially expressed genes in an agricultural important pest, S. litura. PMID:27025647

Genetically modified crops with insect resistance genes from Bacillus thuringiensis Berliner (Bt-plants) are increasingly being cultivated worldwide. Therefore, it is critical to improve our knowledge of their direct or indirect impact not only on target pests but also on non-target arthropods. Hence, this study evaluates comparative leaf consumption and performance of Spodoptera eridania (Cramer), a species that is tolerant of the Cry1Ac protein, fed with Bt soybean, MON 87701×MON 89788 or its near [corrected] non-Bt isoline. Using this species as a model, we assessed [corrected] the comparative performance of the egg parasitoid Telenomus remus Nixon on eggs of S. eridania produced from individuals that fed on these two soybean genotypes [corrected] as larvae. Results showed that Bt soybean did not affect pest foliage consumption, but did reduce larvel duration by two days despite larvae in both treatments having six instars. Nevertheless, survival of S. eridania larvae, pupal weight, sex ratio, fecundity and longevity of female moths, and egg viability did not differ between Bt and non-Bt soybeans. Adult longevity of S. eridania males was increased when caterpillars were fed with Bt soybean versus the near isoline. No adverse effects of this technology were observed for the egg parasitoid T. remus. [corrected].

Bacillus thuringiensis (Bt) insecticidal crystal (Cry) proteins are effective against some insect pests in sprays and transgenic crops, although the evolution of resistance could threaten the long-term efficacy of such Bt use. One strategy to delay resistance to Bt crops is to "pyramid" two or more Bt proteins that bind to distinct receptor proteins within the insect midgut. The most common Bt pyramid in cotton (Gossypium hirsutum L.) employs Cry1Ac with Cry2Ab to target several key lepidopteran pests, including the beet armyworm, Spodoptera exigua (Hübner), which is a serious migratory pest of many vegetable crops and is increasingly important in cotton in China. While cadherin and aminopeptidase-N are key receptors of Cry1 toxins in many lepidopterans including S. exigua, the receptor for Cry2A toxins remains poorly characterized. Here, we show that a heterologous expressed peptide corresponding to cadherin repeat 7 to the membrane proximal extracellular domain (CR7-MPED) in the S. exigua cadherin 1b (SeCad1b) binds Cry1Ac and Cry2Aa. Moreover, SeCad1b transcription was suppressed in S. exigua larvae by oral RNA interference and susceptibility to Cry1Ac and Cry2Aa was significantly reduced. These results indicate that SeCad1b plays important functional roles of both Cry1Ac and Cry2Aa, having major implications for resistance management for S. exigua in Bt crops.

Bacillus thuringiensis strains isolated from Latin American soil samples that showed toxicity against three Spodoptera frugiperda populations from different geographical areas (Mexico, Colombia, and Brazil) were characterized on the basis of their insecticidal activity, crystal morphology, sodium dodecyl sulfate-polyacrylamide gel electrophoresis of parasporal crystals, plasmid profiles, and cry gene content. We found that the different S. frugiperda populations display different susceptibilities to the selected B. thuringiensis strains and also to pure preparations of Cry1B, Cry1C, and Cry1D toxins. Binding assays performed with pure toxin demonstrated that the differences in the toxin binding capacities of these insect populations correlated with the observed differences in susceptibility to the three Cry toxins analyzed. Finally, the genetic variability of the three insect populations was analyzed by random amplification of polymorphic DNA-PCR, which showed significant genetic diversity among the three S. frugiperda populations analyzed. The data presented here show that the genetic variability of S. frugiperda populations should be carefully considered in the development of insect pest control strategies, including the deployment of genetically modified maize in different geographical regions.

Acidic fogs with a pH of 2.0 and duration of 2 hr did not reduce the efficacy of Bacillus thuringiensis var. kurstaki. Therefore, the impact of UV radiation was investigated on the interactions between (1) levels of the antibacterial linear furanocoumarins psoralen, bergapten, and xanthotoxin in Apium graveolens (L.) occurring following a 2.0 pH acidic fog episode, (2) the noctuid Spodoptera exigua, and (3) a sublethal dosage of the microbial pathogen B. thuringiensis var. kurstaki. Mean time to pupation in the absence of UV radiation was significantly extended by the addition of either psoralens or B. thuringiensis. Larvae developing on diets containing B. thuringiensis plus psoralens required nearly 40% longer to pupate than controls, but their effects were additive as the interaction was not significant. Mean time to mortality, a weighted average time of death, was not significantly affected by any of the treatments. In a 2 {times} 2 {times} 2 factorial analysis, all main effects reduced survival significantly, as did the three-way interaction. Thus, antagonistic interactions with psoralens that would reduce the effectiveness of B. thuringiensis in the field were not observed. When pairs of main effects were nested within the two levels of the third factor, several two-way interactions were found. Interestingly, the activity of B. thuringiensis and the psoralens, individually or in combination, was enhanced by exposure to UV radiation. Implications of this research are discussed for both natural and agricultural ecosystems.

The larvicidal activity of 18 phenylpropanoids, 1-18, including phenylpropenoate, phenylpropenal, phenylpropene, and their semisynthetic analogues, were evaluated against the tobacco armyworm, Spodoptera litura (Fab.), to identify promising structures with insecticidal activity. Amongst various phenylpropanoids, isosafrole, a phenylpropene, showed the best activity, with an LC(50) value of 0.6 microg/leaf cm(2), followed by its hydrogenated derivative dihydrosafrole (LC(50)=2.7 microg/leaf cm(2)). The overall larvicidal activity of various phenylpropene derivatives was observed in the following order: isosafrole (6)>dihydrosafrole (16)>safrole (12)>anethole (4)>methyl eugenol (11)>eugenol (13)>beta-asarone (8)>dihydroasarone (18)>dihydroanethole (15). Dihydrosafrole might be a promising compound, although presenting a lower larvicidal activity than isosafrole, because of its better stability and resistance to oxidative degradation (due to the removal of the extremely reactive olefinic bond) in comparison to isosafrole. Such structure-activity relationship studies promote the identification of lead structures from natural sources for the development of larvicidal products against S. litura and related insect pests.

The fall armyworm, Spodoptera frugiperda (J.E. Smith), is one of the chief pests of maize in the Americas. The study of its spatial distribution is fundamental for designing correct control strategies, improving sampling methods, determining actual and potential crop losses, and adopting precise agricultural techniques. In São Paulo state, Brazil, a maize field was sampled at weekly intervals, from germination through harvest, for caterpillar densities, using quadrates. In each of 200 quadrates, 10 plants were sampled per week. Harvest weights were obtained in the field for each quadrate, and ear diameters and lengths were also sampled (15 ears per quadrate) and used to estimate potential productivity of the quadrate. Geostatistical analyses of caterpillar densities showed greatest ranges for small caterpillars when semivariograms were adjusted for a spherical model that showed greatest fit. As the caterpillars developed in the field, their spatial distribution became increasingly random, as shown by a model adjusted to a straight line, indicating a lack of spatial dependence among samples. Harvest weight and ear length followed the spherical model, indicating the existence of spatial variability of the production parameters in the maize field. Geostatistics shows promise for the application of precise methods in the integrated control of pests.

Spodoptera exigua is one of insect causing damage in agriculture sector. This insect can be controlled by a natural biopesticide by combining two agents of biological control, biopesticides Mirabilis jalapa and entomopathogenic fungi Metarhizium anisopliae, considered to be virulent toward a wide range of insects. The objective of research was to determine the effect of biopesticides M. jalapa and fungi M. anisopliae against immune system and mortality of S. exigua. This research used a complete randomized block design with five concentrations Mirabilis jalapa and optimum dose of M. anisopliae. A high dose of M. jalapa (0.8% w/v) is the most effective one to decrease total haemocytes especially granulocyt and plasmatocyt (cellular immune) and decrease the concentration of lectin (humoral immune) from S. exigua (p < 0.05). The combination of M. jalapa (0, 8% w/v) and lethal dose of M. anisopliae 2.59 × 107 spore/ml were significant to increase mortality of S. exigua within 48 hours (p < 0.05).

Spodoptera frugiperda Smith (Lepidoptera: Noctuidae) is a serious pest of corn in several American countries. It is mainly controlled with synthetic insecticides. The objectives of this study were to evaluate the effects of the natural products, neem oil and pyroligneous extract, and the synthetic insecticide, lufenuron, at 2.50 mL water (0.25%) on the mortality of 2-, 4- and 6-day-old caterpillars of S. frugiperda, and their selectivities against fourth instar larvae of Eriopis connnexa Germar (Coleoptera: Coccinellidae). Four- and 6-day-old S. frugiperda caterpillars showed higher mortality after exposure to neem oil (83.33 +/- 0.83 and 89.58 +/- 0.90%, respectively) and lufenuron (95.83 +/- 0.96 and 85.41 +/- 0.83%), compared to pyroligneous extract (68.75 +/- 0.69 and 31.25 +/- 0.31%). The deleterious effect of pyroligneous extract was higher in 2- (83.33 +/- 0.83% mortality) and 4-day-old (68.75 +/- 0.69%) S. frugiperda caterpillars than in 6-day-old caterpillars (31.25 +/- 0.31%). Larval mortality of the predator E. connexa was lower with neem oil and pyroligneous extract (25.00 +/- 0.33%) than with lufenuron (91.66 +/- 1.22%). Neem oil is thus recommended for control of S. frugiperda because of its high toxicity, combined with its relatively low toxicity to larvae of the natural enemy E. connexa.

Cytochrome P450 monooxygenases (P450s) of insects play crucial roles in the metabolism of endogenous and dietary compounds. Tobacco cutworm moth (Spodoptera litura), an important agricultural pest, causes severe yield losses in many crops. In this study, we identified CYP9A40, a novel P450 gene of S. litura, and investigated its expression profile and potential role in detoxification of plant allelochemicals and insecticides. The cDNA contains an open reading frame encoding 529 amino acid residues. CYP9A40 transcripts were found to be accumulated during various development stages of S. litura and were highest in fifth and sixth instar larvae. CYP9A40 was mainly expressed in the midgut and fat body. Larval consumption of xenobiotics, namely plant allelochemicals (quercetin and cinnamic acid) and insecticides (deltamethrin and methoxyfenozide) induced accumulation of CYP9A40 transcripts in the midgut and fat body. Injection of dsCYP9A40 (silencing of CYP9A40 by RNA interference) significantly increased the susceptibility of S. litura larvae to the tested plant allelochemicals and insecticides. These results indicate that CYP9A40 expression in S. litura is related to consumption of xenobiotics and suggest that CYP9A40 is involved in detoxification of these compounds.

We newly sequenced mitochondrial genomes of Spodoptera litura and Cnaphalocrocis medinalis belonging to Lepidoptera to obtain further insight into mitochondrial genome evolution in this group and investigated the influence of optimal strategies on phylogenetic reconstruction of Lepidoptera. Estimation of p-distances of each mitochondrial gene for available taxonomic levels has shown the highest value in ND6, whereas the lowest values in COI and COII at the nucleotide level, suggesting different utility of each gene for different hierarchical group when individual genes are utilized for phylogenetic analysis. Phylogenetic analyses mainly yielded the relationships (((((Bombycoidea + Geometroidea) + Noctuoidea) + Pyraloidea) + Papilionoidea) + Tortricoidea), evidencing the polyphyly of Macrolepidoptera. The Noctuoidea concordantly recovered the familial relationships (((Arctiidae + Lymantriidae) + Noctuidae) + Notodontidae). The tests of optimality strategies, such as exclusion of third codon positions, inclusion of rRNA and tRNA genes, data partitioning, RY recoding approach, and recoding nucleotides into amino acids suggested that the majority of the strategies did not substantially alter phylogenetic topologies or nodal supports, except for the sister relationship between Lycaenidae and Pieridae only in the amino acid dataset, which was in contrast to the sister relationship between Lycaenidae and Nymphalidae in Papilionoidea in the remaining datasets.

Ascoviruses are insect-specific large DNA viruses that mainly infect noctuid larvae, and are transmitted by parasitoids in the fields. Heliothis virescens ascovirus 3h (HvAV-3h) has been recently isolated from Spodoptera exigua, without parasitoid vector identified previously. Here we report that Microplitis similis, a solitary endoparasitoid wasp, could transmit HvAV-3h between S. exigua larvae in the laboratory. When the female parasitoid wasp acquired the virus and served as a vector, the period of virion viability on the ovipositor was 4.1 ± 1.4 days. Infected host larvae were still acceptable for egg laying by parasitoids, and the parasitoids thereafter transmitted virus to healthy hosts. Virus acquisition occurred only from donor hosts between 3 and 9 days post infection. The peak of virus acquisition (80.9 ± 6.3%) was found when M. similis wasps oviposited in larvae that had been inoculated with the virus 7 days previously. When virus infection of the host took place during the life cycle of the parasitoid wasp, it caused 1- to 4-day-old immature parasitoids death in the host, whilst a small proportion of 5- to 6-day-old and the majority of 7-day-old parasitoids larvae survived from the virus-infected hosts. Viral contamination did not reduce the life span or fecundity of female M. similis. PMID:26878829

Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) represents a pest of economic importance in all Western Hemisphere. This polyphagous species has diverged into two populations that have been mainly recognized with various mitochondrial and nuclear molecular markers and named "the rice" and "the corn" strains. In Colombia, both strains have evolved prezygotic and postzygotic isolation. They differ in tolerance to Bacillus thuringiensis (Cry1Ac and Cry1Ab endotoxins) and the insecticides lambda-cyhalothrin and methomyl. In 2014, a wing morphometric analysis made in 159 individuals from a colony showed that both strains significantly differ in wing shape. The species also exhibits sexual dimorphism in the rice strain as in females wing size is larger than in males. Here, we continued this work with another wing morphometric approach in laboratory-reared strains to calculate wing size and shape heritabilities using a full-sib design and in wild populations to determine if this method distinguishes these strains. Our results show that male heritabilities of both traits were higher than female ones. Wild populations were significantly different in wing shape and size. These results suggest that wing morphometrics can be used as an alternative method to molecular markers to differentiate adults from laboratory-reared populations and wild populations of this pest, particularly in males of this species. Finally, Q ST values obtained for wing size and shape further demonstrated that both strains are genetically differentiated in nature.

Two Spodoptera litura glutathione S-transferase cDNAs (Slgste2 and Slgste3) which were cloned from a midgut cDNA, encoded two structurally distinct proteins with a predicted molecular mass of 25 and 24kDa, respectively. Slgste2 and Slgste3 were single-copy genes in the S. litura genome and there was no intron within the genes. The transcripts and proteins of Slgste2 and Slgste3 were predominately present in the midgut of the 5th and 6th instar larvae. The apparent Vmax of the purified SlGSTE2 and SlGSTE3 recombinant proteins towards the substrates glutathione and 1-chloro-2,4-dinitrobenezene (CDNB) were similar. Slgste2 expression was up-regulated by 1-naphthyl methylcarbamate (carbaryl), 1,1,1-trichloro-2,2-bis-(p-chlorophenyl) ethane (DDT), deltamethrin, tebufenozide (RH5992) and Bacillus thuringiensis (Bt), but not affected by malathion, while Slgste3 expression was slightly up-regulated by carbaryl, Bt and DDT, but not affected by RH5992, malathion and deltamethrin. The results suggest that Slgste2 and Slgste3 may play roles in detoxifying various insecticides in S. litura.

Background Oral toxicity of double-stranded RNA (dsRNA) specific to integrin β1 subunit (SeINT) was known in a polyphagous insect pest, Spodoptera exigua. For an application of the dsRNA to control the insect pest, this study prepared a transformed Escherichia coli expressing dsRNA specific to SeINT. Principal Findings The dsRNA expression was driven by T7 RNA polymerase overexpressed by an inducer in the transformed E. coli. The produced dsRNA amount was proportional to the number of the cultured bacteria. The transformed bacteria gave a significant oral toxicity to S. exigua larvae with a significant reduction of the SeINT expression. The resulting insect mortality increased with the fed number of the bacteria. Pretreatment with an ultra-sonication to disrupt bacterial cell wall/membrane significantly increased the insecticidal activity of the transformed bacteria. The larvae treated with the transformed bacteria suffered tissue damage in the midgut epithelium, which exhibited a marked loss of cell-cell contacts and underwent a remarkable cell death. Moreover, these treated larvae became significantly susceptible to a Cry toxin derived from Bacillus thuringiensis (Bt). Conclusions This study provides a novel and highly efficient application technique to use dsRNA specific to an integrin gene by mixing with a biopesticide, Bt. PMID:26171783

Rabbit hemorrhagic disease (RHD) is contagious and highly lethal. Commercial vaccines against RHD are produced from the livers of experimentally infected rabbits. Although several groups have reported that recombinant subunit vaccines against rabbit hemorrhagic disease virus (RHDV) are promising, application of the vaccines has been restricted due to high production costs or low yield. In the present study, we performed codon optimization of the capsid gene to increase the number of preference codons and eliminate rare codons in Spodoptera frugiperda 9 (Sf9) cells. The capsid gene was then subcloned into the pFastBac plasmid, and the recombinant baculoviruses were identified with a plaque assay. As expected, expression of the optimized capsid protein was markedly increased in the Sf9 cells, and the recombinant capsid proteins self-assembled into virus-like particles (VLPs) that were released into the cell supernatant. Rabbits inoculated with the supernatant and the purified VLPs were protected against RHDV challenge. A rapid, specific antibody response against RHDV was detected by an ELISA in all of the experimental groups. In conclusion, this strategy of producing a recombinant subunit vaccine antigen can be used to develop a low-cost, insect cell-derived recombinant subunit vaccine against RHDV.

Expression of a rabbit liver carboxylesterase has been achieved in several different model systems including Escherichia coli, Pichia pastoris, Saccharomyces cerevisiae, Spodoptera frugiperda, and COS7 cells. Although, recombinant protein was observed in E. coli sonicates, little or no enzymatic activity was detected. Similarly, no activity was observed following expression in S. cerevisiae. In contrast, active protein was produced in P. pastoris, from S. frugiperda following baculoviral infection and in COS7 cells following transient transfection of plasmid DNA. For the preparation of small amounts of protein for kinetic and biochemical studies, enzyme expressed in P. pastoris has proved sufficient. However, to produce large amounts of carboxylesterase for structural studies, baculoviral-mediated expression of a secreted form of the protein in S. frugiperda was the most efficient. Using this system, we have generated and purified milligram quantities of essentially pure protein. These results demonstrate that the choice of in vitro system for the generation of large amounts of active carboxylesterase, and probably most endoplasmic reticulum processed proteins, is crucial for high level expression and subsequent purification.

Bacillus thuringiensis vegetative insecticidal proteins (Vip3A) have been recently introduced in important crops as a strategy to delay the emerging resistance to the existing Cry toxins. The mode of action of Vip3A proteins has been studied in Spodoptera frugiperda with the aim of characterizing their binding to the insect midgut. Immunofluorescence histological localization of Vip3Aa in the midgut of intoxicated larvae showed that Vip3Aa bound to the brush border membrane along the entire apical surface. The presence of fluorescence in the cytoplasm of epithelial cells seems to suggest internalization of Vip3Aa or a fragment of it. Successful radiolabeling and optimization of the binding protocol for the 125I-Vip3Aa to S. frugiperda brush border membrane vesicles (BBMV) allowed the determination of binding parameters of Vip3A proteins for the first time. Heterologous competition using Vip3Ad, Vip3Ae, and Vip3Af as competitor proteins showed that they share the same binding site with Vip3Aa. In contrast, when using Cry1Ab and Cry1Ac as competitors, no competitive binding was observed, which makes them appropriate candidates to be used in combination with Vip3A proteins in transgenic crops. PMID:25002420

Bacillus thuringiensis vegetative insecticidal proteins (Vip3A) have been recently introduced in important crops as a strategy to delay the emerging resistance to the existing Cry toxins. The mode of action of Vip3A proteins has been studied in Spodoptera frugiperda with the aim of characterizing their binding to the insect midgut. Immunofluorescence histological localization of Vip3Aa in the midgut of intoxicated larvae showed that Vip3Aa bound to the brush border membrane along the entire apical surface. The presence of fluorescence in the cytoplasm of epithelial cells seems to suggest internalization of Vip3Aa or a fragment of it. Successful radiolabeling and optimization of the binding protocol for the (125)I-Vip3Aa to S. frugiperda brush border membrane vesicles (BBMV) allowed the determination of binding parameters of Vip3A proteins for the first time. Heterologous competition using Vip3Ad, Vip3Ae, and Vip3Af as competitor proteins showed that they share the same binding site with Vip3Aa. In contrast, when using Cry1Ab and Cry1Ac as competitors, no competitive binding was observed, which makes them appropriate candidates to be used in combination with Vip3A proteins in transgenic crops.

Transgenic Agrisure Viptera 3 corn that expresses Cry1Ab, Vip3Aa20, and EPSPS proteins and Agrisure Viptera expressing Vip3Aa20 are used for control of Spodoptera frugiperda (J.E. Smith) and Diatraea saccharalis (F.) in Brazil. To support a resistance management program, resistance risk assessment studies were conducted to characterize the dose expression of Vip3Aa20 protein and level of control against these species. The Vip3Aa20 expression in Agrisure Viptera 3 and Agrisure Viptera decreased from V6 to V10 stage of growth. However, Vip3Aa20 expression in Agrisure Viptera 3 at V6 and V10 stages was 13- and 16-fold greater than Cry1Ab, respectively. The Vip3Aa20 expression in lyophilized tissue of Agrisure Viptera 3 and Agrisure Viptera diluted 25-fold in an artificial diet caused complete larval mortality of S. frugiperda and D. saccharalis. In contrast, lyophilized tissue of Bt11 at the same dilution does not provide complete mortality of these species. Agrisure Viptera 3 and Agrisure Viptera also caused a high level of mortality against S. frugiperda and D. saccharalis. Moreover, 100% mortality was observed for S. frugiperda larvae (neonates through fifth-instar larvae) when fed in corn with the Vip trait technology. Viptera corn achieves a high level of control against S. frugiperda and D. saccharalis providing a high dose, which is an important determination to support the refuge strategy for an effective resistance management program.

The genetically modified maize expressing Vip3Aa20 insecticidal protein from Bacillus thuringiensis Berliner is abiotechnological option for the control of Spodoptera frugiperda (J.E. Smith) and Diatraea saccharalis (F.) in Brazil. To support an Insect Resistance Management program, we conducted studies of baseline susceptibility and monitoring of Brazilian populations of S. frugiperda and D. saccharalis to the Vip3Aa20 insecticidal protein. Neonates were exposed to Vip3Aa20 applied on artificial diet surface. Mortality and growth inhibition were assessed after 7 d. All populations were susceptible to Vip3Aa20. The LC50 ranged from 92.38 to 611.65 ng Vip3Aa20/cm2 for 16 populations of S. frugiperda (6.6-fold variation), and between 61.18 and 367.86 ng Vip3Aa20/cm2 for 6 populations of D. saccharalis (sixfold variation). The EC50 ranged from 21.76 to 70.09 and 48.65 to 163.60 ng Vip3Aa20/cm2 for S. frugiperda and D. saccharalis, respectively. There was a low interpopulation variation in susceptibility to Vip3Aa20, which represents the natural geographic variation in the response, and not the variation caused by previous exposure to selection pressure. For these two pests, the diagnostic concentrations of 2,000 and 3,600 ng of Vip3Aa20/cm2 caused high mortality. These diagnostic concentrations will be used in resistance monitoring programs in Brazil.

Vip proteins, a new group of insecticidal toxins produced by Bacillus thuringiensis, are effective against specific pests including Spodoptera litura. Here, we report construction of a transcriptome database of S. litura by de novo assembly along with detection of the transcriptional response of S. litura larvae to Vip3Aa toxin. In total, 56,498 unigenes with an N50 value of 1,853 bp were obtained. Results of transcriptome abundance showed that Vip3Aa toxin provoked a wide transcriptional response of the S. litura midgut. The differentially expressed genes were enriched for immunity-related, metabolic-related and Bt-related genes. Twenty-nine immunity-related genes, 102 metabolic-related genes and 62 Bt-related genes with differential expression were found. On the basis of transcriptional profiling analysis, we focus on the functional validation of trypsin which potentially participated in the activation of Vip3Aa protoxin. Zymogram analysis indicated that the presence of many proteases, including trypsin, in S. litura larvae midgut. Results of enzymolysis in vitro of Vip3Aa by trypsin, and bioassay and histopathology of the trypsin-digested Vip3Aa toxin showed that trypsin was possibly involved in the Vip3Aa activation. This study provides a transcriptome foundation for the identification and functional validation of the differentially expressed genes in an agricultural important pest, S. litura.

The egg-larval parasitoid Chelonus insularis Cresson is a key parasitoid of the fall armyworm Spodoptera frugiperda (J. E. Smith) during the second field generation in the upper Magdalena River Basin, Colombia. In spite of selection pressure, the higher susceptibility of the second generation fall armyworm larvae to insecticides, compared with that of the first generation, suggests that the parasitism may be responsible for the apparent difference in susceptibility. Parasitized and non-parasitized 2nd-instar larvae of the fall armyworm were tested for susceptibility to chlorpyriphos, methomyl, cypermethrin, and Bacillus thuringiensis in the laboratory, using the leaf dip test. Parasitized larvae were up to 3.93 times more susceptible to chlorpyriphos, 3.71 times to methomyl, and 14.11 times to cypermethrin than non-parasitized larvae. The least effect of parasitism on susceptibility was found for B. thuringiensis. We discuss the negative influence of synthetic insecticide on the parasitoid population dynamics and its impact on insecticide resistance.

The combination of essential oils and Bacillus thuringiensis Berliner may represent an interesting control strategy. Thus, the study tested the following hypothesis: the combination of long pepper oil (Piper hispidinervum L.) and clove (Syzygium aromaticum L.) oils in two concentrations with Xentari WG (Bta) yields a more effective control of Spodoptera frugiperda (JE Smith) (Lepidoptera: Noctuidae) affecting biological and reproductive parameters and leading to changes in the levels of phenoloxidase and nitric oxide in the hemolymph of the pest. The results demonstrate that only long pepper oil, at the highest concentration with Xentari WG (Bta), promotes reduced larval survival. However, both oils with or without the insecticide interfere in the biology and humoral immunity of S.frugiperda. All treatments caused a decrease in the amount of eggs, except for the clove oil at both concentrations without Bta. Therefore, the use of these oils is a promising alternative for the integrated management of S. frugiperda; however, its association with Bta demonstrated no significant increase in their efficiency.

Cell lines derived from the fall armyworm, Spodoptera frugiperda (Sf), are widely used as hosts for recombinant protein production in the baculovirus-insect cell system (BICS). However, it was recently discovered that these cell lines are contaminated with a virus, now known as Sf-rhabdovirus [1]. The detection of this adventitious agent raised a potential safety issue that could adversely impact the BICS as a commercial recombinant protein production platform. Thus, we examined the properties of Sf-RVN, an Sf-rhabdovirus-negative Sf cell line, as a potential alternative host. Nested RT-PCR assays showed Sf-RVN cells had no detectable Sf-rhabdovirus over the course of 60 passages in continuous culture. The general properties of Sf-RVN cells, including their average growth rates, diameters, morphologies, and viabilities after baculovirus infection, were virtually identical to those of Sf9 cells. Baculovirus-infected Sf-RVN and Sf9 cells produced equivalent levels of three recombinant proteins, including an intracellular prokaryotic protein and two secreted eukaryotic glycoproteins, and provided similar N-glycosylation patterns. In fact, except for the absence of Sf-rhabdovirus, the only difference between Sf-RVN and Sf9 cells was SF-RVN produced higher levels of infectious baculovirus progeny. These results show Sf-RVN cells can be used as improved, alternative hosts to circumvent the potential safety hazard associated with the use of Sf-rhabdovirus-contaminated Sf cells for recombinant protein manufacturing with the BICS.

Thousands of Hymenopteran endoparasitoids have developed a unique symbiotic relationship with viruses named polydnavirus (PDVs). These viruses immunocompromise the lepidopteran host allowing the survival of the wasp eggs. In a previous work, we have shown the horizontal transfer of some polydnaviral genes into the genome of the Lepidoptera, Spodoptera exigua. One of these genes, BV2-5 (named gasmin) interferes with actin polymerization, negatively affecting the multiplication of baculovirus in cell culture. In this work, we have focused in the study of the effect of Gasmin expression on different aspects of the baculovirus production. In addition, and since actin polymerization is crucial for phagocytosis, we have studied the effect of Gasmin expression on the larval interaction with bacterial pathogens. Over-expression of Gasmin on hemocytes significantly reduces their capacity to phagocytize the pathogenic bacteria Bacillus thuringiensis. According to these results, gasmin domestication negatively affects baculovirus replication, but increases larvae susceptibility to bacterial infections as pay off. Although the effect of Gasmin on the insect interaction with other pathogens or parasitoids remain unknown, the opposite effects described here could shape the biological history of this species based on the abundance of certain type of pathogens as suggested by the presence of truncated forms of this protein in several regions of the world.

Confined goats spend a substantial part of the day feeding. A poorly designed feeding place increases the risk of feeding in nonphysiological body postures, and even injury. Scientifically validated information on suitable dimensions of feeding places for loose-housed goats is almost absent from the literature. The aim of the present study was, therefore, to determine feeding place dimensions that would allow goats to feed in a species-appropriate, relaxed body posture. A total of 27 goats with a height at the withers of 62 to 80 cm were included in the study. Goats were tested individually in an experimental feeding stall that allowed the height difference between the feed table, the standing area of the forelegs, and a feeding area step (difference in height between forelegs and hind legs) to be varied. The goats accessed the feed table via a palisade feeding barrier. The feed table was equipped with recesses at varying distances to the feeding barrier (5-55 cm in 5-cm steps) at angles of 30°, 60°, 90°, 120°, or 150° (feeding angle), which were filled with the goats' preferred food. In 18 trials, balanced for order across animals, each animal underwent all possible combinations of feeding area step (3 levels: 0, 10, and 20 cm) and of difference in height between feed table and standing area of forelegs (6 levels: 0, 5, 10, 15, 20, and 25 cm). The minimum and maximum reach at which the animals could reach feed on the table with a relaxed body posture was determined for each combination. Statistical analysis was performed using mixed-effects models. The animals were able to feed with a relaxed posture when the feed table was at least 10 cm higher than the standing height of the goats' forelegs. Larger goats achieved smaller minimum reaches and minimum reach increased if the goats' head and neck were angled. Maximum reach increased with increasing height at withers and height of the feed table. The presence of a feeding area step had no influence on minimum and

A retrospective analysis of hand-feeding records and growth data from 3 facilities was performed to determine the growth pattern for 8 toucan species raised in captivity. General philosophies of breeding and rearing were similar but approaches to hand-feeding varied. General hand-feeding and chick management records from hatch to fledging were reviewed for 2 of the 3 facilities. Effective hand-feeding formulas were commercially available and minimally modified. Growth curves were developed. Curves approximated typical expected patterns of nestling growth with no loss of weight at fledging. This study provides a basis for hand-feeding protocols and growth curves to assess development.

Pediatric nurse practitioners are in a key position to help parents learn to effectively feed their children. Classic research studies, current research and clinical observations on feeding have identified children's capabilities, behaviors, and potential competencies with their eating. An appropriate feeding relationship supports children's developmental tasks at every age and allows them to eat the right amount of the proper food to achieve their genetically determined growth endowment. Children must, in turn, attain developmental tasks. In defining and managing feeding problems, the pediatric nurse practitioner may emphasize prevention by building positive feeding interactions, and promoting early detection and management as well as providing treatment for established problems.

Targeting mothers and fathers in breast-feeding promotion programs is recommended as research has found that father's support positively impacts breast-feeding duration and exclusivity. Breast-feeding coparenting refers to the manner in which parents work together to achieve their breast-feeding goals. The Breast-feeding Coparenting Framework was developed on the basis of diverse coparenting models and research related to father's involvement with breast-feeding. This framework consists of 5 components: joint breast-feeding goal setting, shared breast-feeding responsibility, proactive breast-feeding support, father's/partner's parental-child interactions, and productive communication and problem solving. This framework may be of value to policy makers and program providers working to improve breast-feeding outcomes.

In NICU settings, caring for neonates born as early as 23 weeks gestation presents unique challenges for caregivers. Traditionally, preterm infants who are learning to orally feed take a predetermined volume of breast milk or formula at scheduled intervals, regardless of their individual ability to coordinate each feeding. Evidence suggests that this volume-driven feeding model should be replaced with a more individualized, developmentally appropriate practice. Evidence from the literature suggests that preterm infants fed via cue-based feeding reach full oral feeding status faster than their volume-feeding counterparts and have shorter lengths of stay in the hospital. Changing practice to infant-driven or cue-based feedings in the hospital setting requires staff education, documentation, and team-based communication.

A certain attitude and lifestyle is needed if breast feeding is to be successful. It must have priority over all other activities save the very basic ones. The rural environment provides a situation in which breast feeding can flourish . With urbanisation and the accompanying change in lifestyle breast feeding becomes very difficult. Since successful breast feeding needs strong motivation, the change in attitude of urbanised wives makes it even more unlikely. The Singapore experience demonstrates this convincingly; even though awareness of the benefits of breast feeding is widespread, the motivation to breast feed is not. Remedial measures should take into consideration the need to motivate the mother and family, the prevention of commercial infant food companies influencing public opinion, and incentives for the working mothers to breast feed.

Patients who cannot eat enough require alternatives to oral feeding. Tube feeding is one such method for patients with a functioning gut. The techniques for the placement of feeding tubes and diets for tube feeding have improved during the past 20 years. Comfortable thin-bore polyurethane tubes are replacing the thicker vinyl tubes. Long term access routes such as gastrostomy and jejunostomy are being done by endoscopic or radiological techniques. Pre-defined formula feeds have reduced the effort and labour involved in the preparation, storage and administration of blenderized tube feeds. However, the use of soft feeding tubes and commercial formulae will increase the cost of nutrition therapy in India. The ultimate cost-effectiveness of tube feeding will depend on whether it is used appropriately during an illness with adequate delivery of feeds, and whether attention is given to small details by a dedicated nutritional support team. A strong commitment to the scientific use of nutritional support is necessary for getting the best results from tube feeding in everyday practice.

Evidence exists that early life experiences may influence development of characteristic feeding patterns in dairy cattle. The objective of this study was to determine the effect of milk feeding level on performance and development of feeding patterns in dairy calves. Twenty Holstein bull calves were assigned at birth to a milk replacer feeding level, fed via a teat: (1) ad libitum (ADL) or (2) restricted feeding at 5 L/d (0.75 kg of milk replacer) in 2 feedings/d (RES). All calves were offered concentrate ad libitum during the milk feeding stage. Calves were weaned gradually during wk 7 and then fed a pelleted diet ad libitum for 7 wk. Calves were weighed 2 times/wk. Feed and milk intakes were recorded daily. Feeding behavior was recorded from video for 5 d in each of wk 3, 6, 8, 11, and 14. Calves fed ADL consumed 2.6 times more milk, had greater weight gain (1.2 vs. 0.6 kg/d), and consumed less solid feed (45.2 vs. 98.0 g/d) during the milk feeding period than did calves fed RES. As expected, providing milk ADL resulted in more frequent meals (in wk 6, 7.1 vs. 2.0) and more evenly distributed diurnal patterns of feeding activity compared with the RES milk level. When considering all sucking bouts (milk feeding and nonnutritive sucking), no difference was observed between treatments in daily bout frequency or sucking time/bout. Postweaning, calves fed RES had initially greater rate of intake (24.9 vs. 17.8 g/min) and meal size (160.8 vs. 117.2 g/meal). Diurnal feeding patterns also differed in the first week after weaning, with ADL calves feeding less continuously at the time of feed delivery. Meal characteristics and diurnal patterns of feeding activity were similar between calves by the end of the trial. However, milk feeding level had a longer-term effect on variability in feeding behavior, with calves fed RES having greater day-to-day variability in feeding time and meal size. Variability of meal characteristics also decreased over time across treatments

This study aimed to obtain insights into the presence of cadmium, lead, mercury and arsenic in feed materials and feed over time, for the purpose of guiding national monitoring. Data from the Dutch feed monitoring program and from representatives of the feed industry in the period 2007-2013 were used. Data covered the concentrations of cadmium, lead, mercury and arsenic in a variety of feed materials and compound feeds in The Netherlands. Trends in the percentage of samples that exceeded the maximum limit (ML), set by the European Commission, and trends in average, median and 90(th) percentile concentrations of each of these elements per feed material or compound feed were investigated. Based on the results, monitoring for cadmium, lead, mercury and arsenic should focus on feed material of mineral origin, feed material of marine origin, especially fish meal, seaweed and algae as well as feed additives belonging to the functional groups of (i) trace elements (notably cupric sulphate, zinc oxide and manganese oxide for arsenic) and (ii) binders and anti-caking agents. Mycotoxin binders are a new group of feed additives that also need attention. For complementary feed it is important to make a proper distinction between mineral and non-mineral feed because the ML in the latter group is usually lower. In seaweed/algae products a relatively large number of samples contained arsenic concentrations that exceeded the ML. Forage crops in general do not need high priority in monitoring programs, although for arsenic grass meal still needs attention.

A recent resurgence of tuberculosis in the world community has brought the disease into the forefront of communicable disease control. Acknowledging the proven benefits of breast-feeding infants, the question of compatibility arises regarding the safety of breast-feeding an infant in the event of active tuberculosis disease in the mother. This article will discuss the emerging trends of tuberculosis disease and review the evidence that addresses the issues of safety while breast-feeding during tuberculosis treatment.

To isolate antimicrobial-related genes from the beet armyworm, Spodoptera exigua, we performed GeneFishing, a polymerase chain reaction (PCR)-based differential display technique. An attacin-like complementary DNA (cDNA) including a 3'-untranslated region was identified from among 18 over-expressed genes in microbial-infected larvae. The full-length attacin cDNA from S. exigua cDNA (Seattacin) was cloned using rapid amplification of cDNA ends PCR. The attacin-like cDNA transcript was 765 nucleotides in length, and the predicted polypeptide was 254 amino acids in length with a calculated molecular mass of 27.6 kDa and an isoelectric point of 6.44. The protein sequence of the attacin-like cDNA showed high identity to that of Trichoplusia ni (61.2%). The amino acid sequence identity of Seattacin to the orthologous proteins in Bombyx mori, Manduca sexta, Heliothis virescens, Hlicoverpa armigera, Hyphantria cunea, Hyalophora cecropia, and Drosophila melanogaster was 61.2, 46.1, 44.5, 42.2, 39.5, 45.1, and 24.0%, respectively. To examine possible immune functions of the attacin-like cDNA, its expression was investigated by reverse transcriptase PCR analysis after challenging S. exigua with microorganisms. The attacin-like cDNA was expressed at high levels 12 h post-infection, and its expression was slightly induced 4-8 h post-infection compared to control larvae inoculated with sterile water. Furthermore, induced Seattacin showed biological activity against several bacteria including Escherichia coli DH5α, Pseudomonas cichorii, Bacillus subtilis, and Listeria monocytogenes. These results suggest that the attacin-like cDNA of S. exigua codes for antimicrobial peptides.

Evaluations of biological effects of (Pb-CVO) the crude volatile oil of Piper betle leaves on the tobacco cutworm Spodoptera litura were conducted. Pb-CVO was subjected to GC-MS analysis and twenty vital compounds were isolated from the betel leaf oil. Pb-CVO was tested at four different concentrations (0.25, 0.5, 1.0 and 1.5%) against S. litura. The treated insects exhibited dose depended mortality. The mortality rate was significantly higher at the 1.0 and 1.5% Pb-CVO. The LC50 (Lethal concentration) were observed at 0.48% Pb-CVO. Larval and pupal durations increased in all treatment concentrations (0.25, 0.3, 0.4 and 0.5%) whereas, pupal weight decreased compared to control. Adult longevity of S. litura was reduced in all treatments but predominantly in the 0.4 and 0.5% Pb-CVO. Correspondingly, mean fecundity rate was reduced at all concentrations compared to control. Histological studies of larvae mid-gut profiles of S. litura were severely damaged in 1.0 and 1.5% and showed abnormalities in mid-gut cells with 0.25 and 0.5% Pb-CVO treatments. Earthworm toxicity illustrated that 0.1% of chemical insecticides (monocrotophos and cypermethrin) varied widely in their contact toxicities compared to 0.5 and 1.0% Pb-CVO and control in both contact filter paper and artificial soil test. These findings suggest that twenty essential compounds of betel leaf oil were significant inhibitors of the development and caused behavioral changes of S. litura. Treatment with betel leaf oil at these concentrations had no adverse effect on earthworm populations.

The human alpha-3/4-fucosyltransferase III (Fuc-TIII) participates in the synthesis of Lewis determinants. The enzyme from human sources is scarce and heterogeneous. In this paper we describe the expression of a secreted form of Fuc-TIII (SFT3) in two insect cell lines, Spodoptera frugiperda (Sf9) and Trichoplusia ni (Tn), using the baculovirus expression system. The Sf9 cells secreted approx. 0.4 unit/l (1 mg/l) of the enzyme. The Tn cells secreted approx. 3-fold this amount. A large proportion of active protein was accumulated in the two cell lines (50 and 75% respectively for Sf9 and Tn cells, on the fourth day after infection) indicating a possible limitation not only of the folding machinery, but also a saturation of the secretory pathway. SFT3 was purified by cation-exchange chromatography followed by affinity chromatography. The enzyme from the Tn cell line had a lower global charge, possibly due to post-translational modifications, such as phosphorylation or sulphation. The two glycosylation sites from SFT3 were occupied. SFT3 secreted by Sf9 cells was completely deglycosylated by peptide-N-glycanase F, whereas 50% of SFT3 secreted by Tn cells was resistant to deglycosylation by this enzyme. The apparent kinetic parameters determined with the type I acceptor were k(cat)=0.4 s(-1) and K(m)=0.87 mM for the SFT3 secreted by Tn cells, and k(cat)=0.09 s(-1) and K(m)=0.76 mM for the SFT3 secreted by Sf9 cells, indicating that the enzymes had substrate affinities within the same order of magnitude as their mammalian counterpart. Furthermore, SFT3 secreted by either cell type showed a clear preference for type 1 carbohydrate acceptors, similarly to human Fuc-TIII. PMID:11171070

Insect cytochrome P450 monooxygenases (CYPs or P450s) play an important role in detoxifying insecticides leading to resistance in insect populations. A polyphagous pest, Spodoptera litura, has developed resistance to a wide range of insecticides. In the present study, a novel P450 gene, CYP321B1, was cloned from S. litura. The function of CYP321B1 was assessed using RNA interference (RNAi) and monitoring resistance levels for three commonly used insecticides, including chlorpyrifos, β-cypermethrin and methomyl. The full-length complementary DNA sequence of CYP321B1 is 1814 bp long with an open reading frame of 1 488 bp encoding 495 amino acid residues. Quantitative reverse-transcriptase polymerase chain reaction analyses during larval and pupal development indicated that CYP321B1 expression was highest in the midgut of fifth-instar larvae, followed by fat body and cuticle. The expression of CYP321B1 in the midgut was up-regulated by chlorpyrifos, β-cypermethrin and methomyl with both lethal concentration at 15% (LC15 ) (50, 100 and 150 μg/mL, respectively) and 50%(LC50 ) dosages (100, 200 and 300 μg/mL, respectively). Addition of piperonyl butoxide (PBO) significantly increased the toxicity of chlorpyrifos, β-cypermethrin and methomyl to S. litura, suggesting a marked synergism of the three insecticides with PBO and P450-mediated detoxification. RNAi-mediated silencing of CYP321B1 further increased mortality by 25.6% and 38.9% when the fifth-instar larvae were exposed to chlorpyrifos and β-cypermethrin, respectively, at the LC50 dose levels. The results demonstrate that CYP321B1 might play an important role in chlorpyrifos and β-cypermethrin detoxification in S. litura.

Gossypium hirsutum (commercial cooton) is one of the most economically important fibers sources and a commodity crop highly affected by insect pests and pathogens. Several transgenic approaches have been developed to improve cotton resistance to insect pests, through the transgenic expression of different factors, including Cry toxins, proteinase inhibitors, and toxic peptides, among others. In the present study, we developed transgenic cotton plants by fertilized floral buds injection (through the pollen-tube pathway technique) using an DNA expression cassette harboring the cry1Ia12 gene, driven by CaMV35S promoter. The T0 transgenic cotton plants were initially selected with kanamycin and posteriorly characterized by PCR and Southern blot experiments to confirm the genetic transformation. Western blot and ELISA assays indicated the transgenic cotton plants with higher Cry1Ia12 protein expression levels to be further tested in the control of two major G. hirsutum insect pests. Bioassays with T1 plants revealed the Cry1Ia12 protein toxicity on Spodoptera frugiperda larvae, as evidenced by mortality up to 40% and a significant delay in the development of the target insects compared to untransformed controls (up to 30-fold). Also, an important reduction of Anthonomus grandis emerging adults (up to 60%) was observed when the insect larvae were fed on T1 floral buds. All the larvae and adult insect survivors on the transgenic lines were weaker and significantly smaller compared to the non-transformed plants. Therefore, this study provides GM cotton plant with simultaneous resistance against the Lepidopteran (S. frugiperda), and the Coleopteran (A. grandis) insect orders, and all data suggested that the Cry1Ia12 toxin could effectively enhance the cotton transgenic plants resistance to both insect pests. PMID:26925081

In Brazil, Spodoptera frugiperda (J. E. Smith) and Chrysodeixis includens (Walker) are important cotton pests and target of control of Bollgard II (Cry1Ac/Cry2Ab2) and WideStrike (Cry1Ac/Cry1F) cotton technologies. To subsidize an insect resistance management program, we conducted laboratory studies to evaluate the toxicity of these Bt cotton plants throughout larval development of S. frugiperda and C. includens. In bioassays with leaf disc, the efficacy of both Bt cotton plants against neonates was >80% for S. frugiperda and 100% for C. includens. However, S. frugiperda larvae that survived on Bt cotton had >76% of growth inhibition and stunting. In bioassays with S. frugiperda and C. includens larvae fed on non-Bt near-isoline during different time period (from 3 to 18 d) and then transferred to Bollgard II or WideStrike leaves showed that larval susceptibility decreased as larval age increased. For Bollgard II cotton, in all S. frugiperda instars, there were larvae that reached the pupal and adult stages. In contrast, on WideStrike cotton, a few larvae in fifth and sixth instar completed the biological cycle. For C. includens, some larvae in sixth instar originated adults in both Bt cotton plants. In conclusion, Bollgard II and WideStrike cotton technologies showed high efficacy against neonates of S. frugiperda and C. includens. However, the mortality of these species decreases as larval age increase, allowing insect survival in a possible seed mixture environment and favoring the resistance evolution.

Transgenic crops producing Bacillus thuringiensis (Bt) proteins have become a primary tool in pest management. Due to the intensive use of Bt crops, resistance of the fall armyworm, Spodoptera frugiperda, to Cry1F maize has occurred in Puerto Rico, Brazil, and some areas of the southeastern U.S. The sustainability of Bt crops faces a great challenge because the Cry1F-maize resistant S. frugiperda may also infest other Bt crops in multiple cropping ecosystems. Here we examined the survival and plant injury of a S. frugiperda population selected with Cry1F maize on three single-gene and five pyramided Bt cotton products. Larvae of Cry1F-susceptible (SS), -heterozygous (RS), and -resistant (RR) genotypes of S. frugiperda were all susceptible to the pyramided cotton containing Cry1Ac/Cry2Ab, Cry1Ac/Cry1F/Vip3A, Cry1Ab/Cry2Ae, or Cry1Ab/Cry2Ae/Vip3A, and the single-gene Cry2Ae cotton. Pyramided cotton containing Cry1Ac/Cry1F was effective against SS and RS, but not for RR. These findings show that the Cry1F-maize selected S. frugiperda can cause cross-crop resistance to other Bt crops expressing similar insecticidal proteins. Resistance management and pest management programs that utilize diversify mortality factors must be implemented to ensure the sustainability of Bt crops. This is especially important in areas where resistance to single-gene Bt crops is already widespread. PMID:27301612

Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae) is an important pest of several crops in the western hemisphere. This insect has genetically differentiated into two host-associated populations: the corn (Zea mays L.) and the rice (Oryza sativa L.) strains. The corn strain also is found in cotton (Gossypium hirsutum L.) and sorghum and the rice strain in Bermuda grass [Cynodon dactylon (L.) Pers.] and millet. In the United States and Brazil, lines from corn, rice, Bermuda grass, and millet were used to evaluate the resistance of both strains to various insecticides, and found that the corn strain is more resistant than the rice strain. However, in these studies the larvae were not genotyped. In Colombia, genotyping of fall armyworm is necessary because the rice strain also can be found in corn fields. In this work, collected larvae from corn and rice fields from Tolima (central Colombia) were genotyped and evaluated for the resistance to methomyl and lambda-cyhalothrin. We found that the rice strain does not significantly differ in resistance to methomyl compared with the corn strain but it develops tolerance more rapidly to lambda-cyhalothrin. The eggs viability of treated females also was significantly affected by methomyl on each generation. The realized heritability of resistance was higher for lambda-cyhalothrin (0.23-0.42) than for methomyl (0.04-0.14). The number of generations needed for 10-fold increase in resistance is approximately 11.5 generations for methomyl and 6.5 for lambda-cyhalothrin. Finally, the genetic basis of resistance to both insecticides involves few recessive autosomal genes. The results obtained here suggest that methomyl is a better option than lambda-cyalothin to control fall armyworm.

Rhinacanthus nasutus (Acanthaceae) is a shrub reported to contain insecticidal activities. The current study was conducted to determine whether R. nasutus constituents could inhibit benzyloxyresorufin O-debenzylation (BROD) mediated by CYP6AA3 and CYP6P7. Both enzymes have shown pyrethroid degradation activity and been implicated to play role in pyrethroid resistance in Anopheles minumus (Theobald) mosquito, a malaria vector. Three compounds, rhinacanthin-A, -B, and -C that exhibited potent inhibitory activity were isolated and purified from aerial part of R. nasutus. Their kinetic parameters and modes of inhibition were determined. Rhinacanthin-B was the most potent inhibitor in in vitro inhibition assay and exhibited mechanism-based inhibition against both CYP6AA3 and CYP6P7. Rhinacanthin-C which is a major compound of R. nasutus reversibly inhibited both enzymes in vitro with 2-4 folds less inhibitory potency than rhinacanthin-B. In contrast, rhinacanthin-A reversibly inhibited CYP6AA3, but inhibition against CYP6P7 was a mechanism-based inhibition type. Where mechanism-based inhibition was found, the inhibition showed characteristic of time-, concentration-dependence, and requirement of NADPH. Using 3-(4, 5-dimethylthiazol-2-y-l)-2, 5-diphenyltetrazolium bromide (MTT) cytotoxicity assay in intact Spodoptera frugiperda (Sf9) cells, the three compounds increased susceptibility of CYP6AA3- and CYP6P7-expressing cells to cypermethrin cytotoxicity because of inhibition effect on mosquito enzymes. The combined inhibition effect on mosquito cytochrome P450 enzyme and synergistic effect on cypermethrin cytotoxicity of the three R. nasutus compounds could be beneficial for resistance management strategies in mosquito vector control.

Heat shock proteins (Hsps) are important chaperones, which are involved in various signal pathways and regulate lots of physiological processes. Early research suggested that some Hsps are involved in insect development. However, few studies have been carried out to explore the roles of Hsps, especially in larval-pupal metamorphosis. In the present study, 49 Hsp unigenes were identified in the Spodoptera litura transcriptome and their mRNA expression profiles during midgut metamorphosis were examined using a tag-based digital gene expression system. The genes with the most different levels of expression were then cloned and their expression patterns in midguts from sixth instar larvae to pupae were analysed using real time quantitative PCR. The responses of these genes to juvenile hormone (JH) and 20-hydroxyecdysone (20E) were also studied. The results showed that the mRNA levels of 22 Hsp unigenes changed significantly during midgut metamorphosis. Amongst these 22 unigenes, hsp70, hsp20.4 and hsp20.8 were the most up-regulated members, and hsp15.9, hsp19.3 and hsp22.0 were the most down-regulated ones. Further studies showed that hsp70, hsp20.4 and hsp20.8 were remarkably up-regulated by JH. In addition, 20E slightly increased the mRNA levels of both hsp20.4 and hsp20.8. However, hsp15.9, hsp19.3 and hsp22.0 did not respond to either JH or 20E. These results indicate that Hsp70 and small Hsps (sHsps) are probably the major players in midgut metamorphosis in S. litura. The current findings provide valuable insights into the roles of the Hsp superfamily in insect metamorphosis.

Isoflavonoids are a characteristic family of natural products in legumes known to mediate a range of plant-biotic interactions. For example, in soybean (Glycine max: Fabaceae) multiple isoflavones are induced and accumulate in leaves following attack by Spodoptera litura (Lepidoptera: Noctuidae) larvae. To quantitatively examine patterns of activated de novo biosynthesis, soybean (Var. Enrei) leaves were treated with a combination of plant defense elicitors present in S. litura gut content extracts and L-α-[(13)C9, (15)N]phenylalanine as a traceable isoflavonoid precursor. Combined treatments promoted significant increases in (13)C-labeled isoflavone aglycones (daidzein, formononetin, and genistein), (13)C-labeled isoflavone 7-O-glucosides (daidzin, ononin, and genistin), and (13)C-labeled isoflavone 7-O-(6″-O-malonyl-β-glucosides) (malonyldaidzin, malonylononin, and malonylgenistin). In contrast levels of (13)C-labeled flavones and flavonol (4',7-dihydroxyflavone, kaempferol, and apigenin) were not significantly altered. Curiously, application of fatty acid-amino acid conjugate (FAC) elicitors present in S. litura gut contents, namely N-linolenoyl-L-glutamine and N-linoleoyl-L-glutamine, both promoted the induced accumulation of isoflavone 7-O-glucosides and isoflavone 7-O-(6″-O-malonyl-β-glucosides), but not isoflavone aglycones in the leaves. These results demonstrate that at least two separate reactions are involved in elicitor-induced soybean leaf responses to the S. litura gut contents: one is the de novo biosynthesis of isoflavone conjugates induced by FACs, and the other is the hydrolysis of the isoflavone conjugates to yield isoflavone aglycones. Gut content extracts alone displayed no hydrolytic activity. The quantitative analysis of isoflavone de novo biosynthesis, with respect to both aglycones and conjugates, affords a useful bioassay system for the discovery of additional plant defense elicitor(s) in S. litura gut contents that specifically

To test our hypothesis that substitution of domain III of Bacillus thuringiensis delta-endotoxin (Cry) proteins might improve toxicity to pest insects, e.g., Spodoptera exigua, in vivo recombination was used to produce a number of cryIA(b)-cryIC hybrid genes. A rapid screening assay was subsequently exploited to select hybrid genes encoding soluble protoxins. Screening of 120 recombinants yielded two different hybrid genes encoding soluble proteins with domains I and II of CryIA(b) and domain III of CryIC. These proteins differed by only one amino acid residue. Both hybrid protoxins gave a protease-resistant toxin upon in vitro activation by trypsin. Bioassays showed that one of these CryIA(b)-CryIC hybrid proteins (H04) was highly toxic to S. exigua compared with the parental CryIA(b) protein and significantly more toxic than CryIC. In semiquantitative binding studies with biotin-labelled toxins and intact brush border membrane vesicles of S. exigua, this domain III substitution appeared not to affect binding-site specificity. However, binding to a 200-kDa protein by CryIA(b) in preparations of solubilized and blotted brush border membrane vesicle proteins was completely abolished by the domain III substitution. A reciprocal hybrid containing domains I and II of CryIC and domain III of CryIA(b) did bind to the 200-kDa protein, confirming that domain III of CryIA(b) was essential for this reaction. These results show that domain III of CryIC protein plays an important role in the level of toxicity to S. exigua, that substitution of domain III may be a powerful tool to increase the repertoire of available active toxins for pest insects, and that domain III is involved in binding to gut epithelium membrane proteins of S. exigua. PMID:8633853

A complementary DNA encoding a TIA-1-type RNA-binding protein (SfTRN-1) was isolated from cultured cells of the fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae), to characterize its function. The deduced 388-amino acid sequence of SfTRN-1, which possessed three RNA recognition motifs (RRMs) followed by a C-terminal auxiliary domain, showed significant homology with mammalian TIA-1/TIAR and silkworm BmTRN-1, factors important in the metabolism of transcripts. It was found that inhibition of SfTRN-1 gene expression by a transfected oligonucleotide encoding the antisense sequence led to a marked increase in the production of a reporter protein and the amount of reporter transcript in the cultured cells. In addition, overexpression of the recombinant full-length SfTRN-1 open reading frame in the cultured cells led to a decrease in reporter protein production, but the truncated RRM1-3 domain lacking the C-terminal auxiliary domain lost its activity. Analysis using a GFP-fused recombinant protein revealed that, unlike mammalian TIA-1/TIAR, SfTRN-1, most likely shuttling between the nucleus and cytoplasm, had the characteristic of being largely distributed in the cytoplasm, where it perhaps acts to reduce the amount of transcripts, and that RRM1 and RRM3 were related to its nuclear accumulation, but RRM2 to its nuclear export. Furthermore, the posterior half of the auxiliary domain was also found to be related to its nuclear export. This study indicates that respective RRM subdomains of SfTRN-1 play distinct roles important to its subcellular distribution, and it identified unique systems for the distribution and functional regulation of the TIA-1 family in insect cells, ones which are clearly different from those in mammalian cells.

It is challenging to find genes with stable transcripts for use as reference genes for quantitative realtime polymerase chain reaction (qRT-PCR) during viral infection. Autographa californica nucleopolyhedrovirus (AcMNPV) is known to globally shut off host gene transcription in Sf21 cells and to modify their cytoskeletons. In this study, seven host genes were selected for validation as references for gene expression experiments using qRT-PCR. Two of them, ecdysoneless (ECD) and myosin showed stable RNA levels in our previous microarray study at 6, 12, and 24 hpi for both genes and 48 hpi for ECD. The others, actin, tubulin, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and 28S ribosome (28S), are commonly employed as reference genes for qRT-PCR. Ribosomal protein L35 (L35) gene was selected to test if ribosomal protein genes show stable RNA transcript levels similar to 28S and 18S rRNA and to validate the microarray data. In addition to 28S, previously known to have stable transcript levels, qRT-PCR showed that ECD transcript levels remained constant throughout the time course of AcMNPV infection. Transcripts of cytoskeleton genes such as actin, tubulin, and myosin declined dramatically as the infection progressed. GAPDH and L35 transcripts also declined over time. These results indicate that ECD is a reliable reference gene for qRT-PCR experiments during AcMNPV infection of Spodoptera frugiperda cells. Although 28S could be used as a reference gene for these experiments, it is less useful than ECD because of its abundance, which might make it difficult to establish an accurate baseline value for data analysis.

Several mutants of the Bacillus thuringiensis Cry1Ca toxin affected with regard to specific activity towards Spodoptera exigua were studied. Alanine was used to replace single residues in loops 2 and 3 of domain II (mutant pPB19) and to replace residues 541–544 in domain III (mutant pPB20). Additionally, a Cry1Ca mutant combining all mutations was constructed (mutant pPB21). Toxicity assays showed a marked decrease in toxicity against S. exigua for all mutants, while they retained their activity against Manduca sexta, confirming the importance of these residues in determining insect specificity. Parameters for binding to the specific receptors in BBMV (brush border membrane vesicles) of S. exigua were determined for all toxins. Compared with Cry1Ca, the affinity of mutant pPB19 was slightly affected (2-fold lower), whereas the affinity of the mutants with an altered domain III (pPB20 and pPB21) was approx. 8-fold lower. Activation of Cry1Ca protoxin by incubation with S. exigua or M. sexta BBMV revealed the transient formation of an oligomeric form of Cry1Ca. The presence of this oligomeric form was tested in the activation of the different Cry1Ca mutants, and we found that those mutated in domain II (pPB19 and pPB21) could not generate the oligomeric form when activated by S. exigua BBMV. In contrast, when oligomerization was tested using BBMV prepared from M. sexta, all of the Cry1Ca mutants showed the formation of a similar oligomeric form as did the wild-type toxin. Our results show how modification of insect specificity can be achieved by manipulation of different parts of the toxin structure involved in different steps of the mode of action of B. thuringiensis toxins. PMID:15320864

The effects of sublethal concentrations 0.1, 0.5, and 1.2 µg mL-1of the chitin synthesis inhibitor, hexaflumuron, on larval growth and development, the count and proportion of hemocytes, and carbohydrate content (trehalose and glyceride) in hemolymph were investigated in the cutworm, Spodoptera litura (Fabricious) (Lepidoptera: Noctuidae). When 3rdinstar larvae were subjected to the sublethal concentrations, there were dose-dependent effects on larval weight and length of each instar larvae, percent pupation and the duration of development. Most of the larvae died during the molting process at all concentrations. Few individuals from 0.5 and 1.2 µg mL -1concentrations could develop to the 6thinstar, while the pupae emerging from the 0.1 µg mL -1concentrations did not exceed 16% of the number of the initial larvae. In 5thinstar S. litura, the total number of hemocytes was significantly increased at 24 hours post—treatment, whereas the proliferation of hemocytes was inhibited, plasmatocyte pseudopodia contracted, and granulocyte expanded at 96 hours post—treatment. The increases of plasmatocyte count and the decreases of granulocyte count were dose—dependent. The longer treatment time of the sublethal concentrations increased the content of total carbohydrate and trehalose in hematoplasma, and was dose—dependent in hemocytes. The content of glyceride in hemolymph was significantly higher at 24 hours post—treatment, but gradually returned to normal levels at 96 hours post—treatment as compared with the control. The results suggested that sublethal concentrations of hexaflumuron reduced S. litura larval survival and interfered with hemolymph physiological balances. PMID:22958164